eChapters

Two types of molecular weight (MW) standards are routinely used. The Lambda/HindIII and PhiX174/HaeIII MW standards provide a useful reference for calculating molecular weights of large and small DNA fragments, respectively, after electrophoretic separation; the “internal MW standards” provide a means for normalizing fragment migration distances within each lane to facilitate comparisons between lanes on the same or different luminographs in fingerprinting studies.

Electrophoresis is an analytical method frequently used in molecular biology and medicine. It is applied for the separation and characterization of proteins, nucleic acids and subcellular-sized particles like viruses and small organelles. Its principle is that the charged particles of a sample migrate in an applied electrical field. An electric current is passed through a medium containing the molecules, and each molecule travels at a different rate depending on its electrical charge, size and shape. Separation by electrophoresis is based on these differences. In electrophoresis, agarose and acrylamide gels are used for electrophoresis of nucleic acids and proteins.

What is Gel Electrophoresis?

A gel is a solid form. The term electrophoresis describes the migration of charged particle under the influence of an electric field. Electro refers to the energy of electricity. Phoresis, from the Greek verb phoros, means “to carry across.” Thus, gel electrophoresis refers to the technique in which molecules are forced across a span of gel, motivated by an electrical current. Activated electrodes at either end of the gel provide the driving force. A molecule’s properties determine how rapidly an electric field can move the molecule through a gelatinous medium.

Although it can be argued that due to its overall trendiness, they have been a lot of recent developments in making it more fun to work with. As a molecule, it is very similar to DNA (the most notable chemical difference is its flexibility), but suffers because the nucleases (RNases) that can destroy it are notoriously very stable and very difficult to get rid of.

What is DNA

DNA is a simple molecule

DNA is negatively charged

DNA has lots of ring structures; ring structures are generally quite hydrophobic.

DNA is a very robust molecule.

DNA work has lots of toys.

1. pH meter

pH meter/electrode systems are the most common method of measuring pH in the biology laboratory. Although more expensive and more complicated to use than chemical indicators, pH meters provide greater accuracy, sensitivity, and flexibility. When used properly, pH meters can measure the pH of a solution to the nearest 0.1 pH unit or better, and they can be used with a variety of samples.

Abstract

Information and Communication Technology (ICT) played and continue to play pivotal role in better decision making of farmers for plant health management under the climate change scenario. For judicious plant health management by farmers through the use of ICT tools (viz. interactive Kiosk, Mobile Apps, Whatsapp, Webportal), information need and information access network to be assessed. Based on farmers’ information need and network, information delivery tree should be designed in ICT tools. Under this backdrop this research work was carried out among the farmers of Bhagalpur district of Bihar. The sample size was 120. From the study it was noted that the major information needs of the farmers were seed availability, crop disease management, insect pest management, weed management and weather information and pesticide application. Farmers were enthusiastic to improve their knowledge on insect pests and disease identification and management. Specifically, farmers need information on insect pest management of wheat (aphid), maize (stem borer), vegetables (fruit borer), pulses (pod borer) and oilseeds (aphid) and also disease management of wheat (leaf blight and spot blotch disease), maize (Turcicum leaf blight), vegetables (wilt and leaf spot) and pulses (wilt disease). Farmers decision making was positively and significantly correlated with crop disease management(0.196*),insect pest management of crop(0.208*) and weed management(0.239**). Through the Path Analysis it was noted that farmers decision making on disease management was mediated by the variables - Bihar Agricultural University-Kisan Diary, Mobile Apps(Bihar Krishi Apps) and Kisan Salahkar. So, it can be concluded that these much needed information should be provided to the farmers though different ICT tools and extension personnel should be trained to operate these ICT tools for accelerated use of ICT tools by the farmers.

Abstract

Bacterial diseases of solanaceous crops like tomato, brinjal, chilli, capsicum and potato are the most serious and destructive diseases affecting both under field and greenhouse conditions. Under moist field conditions they can cause epidemics to affect total crop loss, quality of the produce and reduce market value. These diseases can be distinguished from each other on the basis of their symptoms appearance of the various parts of the plant as well as by using advanced techniques of diagnosis. Diagnosis of the disease is important step to detect pathogen at early stage of disease and subsequently manage it effectively in efficient manner.

Keywords: Bacterial diseases, solanaceous crops, symptoms, diagnosis.

Introduction

Bacterial diseases of solanaceous crops like tomato, brinjal, chilli, capsicum and potato are the most serious and destructive diseases affecting both under field and greenhouse conditions. Under moist field conditions they can cause epidemics to affect total crop loss, quality of the produce and reduce market value. These diseases can be distinguished from each other on the basis of their symptoms appearanceofthe various parts of the plant as well as by using advanced techniques of diagnosis. Diagnosis of the disease is important step to detect pathogen at early stage of disease and subsequently manage it effectively in efficient manner. Therefore, the early detection of the bacterium from asymptomatic plants, soil, and water and plant residues could facilitate elimination and certainly reduce the risk of crop loss. Powerful tools for the identification and detection of the bacterium in diverse substrates (plant, seed, water and soil) are available for diagnosis of the bacterial diseases. DNA amplification of pathogen offers many advantages over classical techniques; neither purification nor cultivation of the pathogen is required and the specificity, sensitivity and response time of tests are improved. Nevertheless, the PCR method has not yet become a routine diagnostic tool for many laboratories, mainly because of the inhibition of the amplification reaction by compounds contained in crude bacterial extracts, which give false negative results or low detection sensitivity.

Abstract

Onion (Allium cepa L.) and garlic (Allium sativum L.) are important members of the family Amaryllidaceae. Bulbs, which are the edible component in both onion and garlic grow underground and are of nutritional and medicinal importance. These underground bulb vegetables are affected by many diseases caused by different microorganisms like fungi, bacteria, nematode and viruses causing huge economic loss. Stemphylium leaf blight caused by Stemphylium vesicarium, is the major fungal disease affecting their productivity of the edible bulbs adversely.The fungus primarily attacks the leaves but can affect bulbs in storage. Initial infection is characterised by appearance of small greyish white oval spots on the leaves. These spots are often surrounded by a yellowish halo with irregular margins. Fungal pseudothecia may over-winter in contaminated plant material or may survive in the soil having plant debris and ascospores are the primary inoculums in the following season. Spores require rain or persistent dew to cause infection. This disease can spread rapidly when environmental conditions are favourable for spore development. Suitable integration of more efficient treatments like fungicides, bio agents in combination with appropriate cultural control provide better management of the Stemphylium blight disease

Abstract

India ranks first among world’s mango producing countries accounting for about 50 % of the world’s total mango production. In Bihar it is the most important fruit crop as well as occupies a large area affecting socio economic conditions of the region. The crop suffers from over 140 diseases all over the world. From young seedlings to mature trees and fruits in their all stages from start to storage may suffer from a number of biotic and abiotic stresses. All parts of the plant namely, trunk, branch, twigs, leaf, petiole, flower and fruits are prone to attack by a number of pathogens The changes in climatic patterns and shifting towards commercial cultivation of this crop has seen the emergence of some diseases which can cause huge losses to the production of mango over the years. One such problem, decline in mango is reported to be a complex problem involving multitude of factors. Decline disorders are recognized in virtually all mango-producing regions of the world. In most cases, the disease has been characterized by leaf drooping and drying; bark splitting, gum secretion from branches and main trunk, bud necrosis, dieback, vascular discoloration and death of the whole tree. The bark beetle Hypocryphalus mangiferae is closely associated with trees affected by mango sudden decline disease. Report suggests the causal agent to be Botryodiplodia theobromae, Ceratocystis manginecans, Fusarium spp, Colletotrichum sp, Pestalosiopsis maniferae. In most of the cases Botryodiplodia theobromae, Ceratocystis spp are associated with it. Although, none of the pathogens isolated could singly incite the complete decline symptoms under artificial inoculation on mango seedlings indicating towards complex etiology of the disease. Since pathogens are endophytic in nature; mangement of the pathogen after infection is difficult and relies more upon prohytalctic measures. Over all plant health, its nutritional status and abiotic conditions of flood and drought are also the pre disposing factors to decline in Mango. Effective management of decline in mango needs holistic approaches using optimum nutrition, general sanitation, routine crop protection schedule, weed management and appropriate intercropping practices.

Abstract

Fungal rot and wilt caused by Sclerotinia sclerotiorum is a devastating disease of vegetables causing huge loss. The fungus has a broad host of more than 400 species including diverse vegetable crops, viz., carrot, potato, tomato, green pepper, eggplant, lettuce, peas and beans, cole crops, bottle gourd, pointed gourd, etc. Being able to overwinter in the soil in form of sclerotia upto 5 years and having such a wide host range, management of the disease becomes very difficult. Thus development of resistant varieties can be the only long term strategy for control of the pathogen. The screening of the existing genotypes and identification of resistant genotypes forms the base of any resistant breeding programme. The widely available genotypes of different crops have been screened by several researchers and resistant genotypes have been identified in cauliflower, vegetable soybean, peas, tomato, etc. These genotypes are further used in breeding programmes for as donors for resistance to Sclerotinia sclerotiorum.

Keywords: Vegetables, Sclerotinia, Resistance breeding, Genotypic screening

Abstract

Palms are generally grown commercially in tropical climate. But some of the palms like coconut, areca nut and palmyra palm are cultivated successfully in West Bengal, Orissa, Assam, and some pockets of Bihar that lies under sub tropics of India.Due to unsuitable climatic factor their production is more affected by abiotic and biotic stress including different kinds of diseases. The important disease are bud rot, basal stem end rot, nut shedding,stem bleeding in coconut, mahali disease, foot rot, stem breaking in arecanut and palmyra palm in the region have negligible incidence of diseases. These destructive diseases are the major constrain in production and causes severe yield loss in these palm fruits. Therefore effective management strategies are required by continuous monitoring the incidence and severity of these diseases caused by different pathogens.

Integrated disease management is the most accepted strategies for sustainable crop production that involves the holistic approach integrating cultural, physical, mechanical, chemical and biological means to control the disease. However, results of the disease management trials depend upon soil condition, age of the crop, stage of disease development and weather parameters of that particular area. Although it is impossible to manage a field free of pathogen but considerable reduction of the disease can be achieved through a proper integrated disease management system. Thus the causes and remedy of the diseases prevalent mainly in palms particularly in subtropical conditions has been reviewed for better management and sustainable production.

Introduction

The term biological control was first used in scientific literature in 1914. Baker and Cook (1974) described biological control as “reduction of inoculum density or disease producing activities of a pathogen or parasite in its active or dormant state, by one or more organisms, accomplished naturally or thoroughly manipulation of environment, host, or antagonists or by mass introduction of one or more antagonists.”

Certain genera of fungi have greater potential for being effective bio-control agents eg. Trichoderma, Gliocladium, Chaetomium; entomogenous fungi as Entomophthora, Beauveria, Metarrhizium, Hirsutella; Nematophagous viz., Catenaria, Zoopage, Stylopage, Paecilomyces and predaceaus fungi like Arthrobotrys, Dictylella, Dactylaria and Nematoctonus (Narain, 2018). Biopesticide formulations based on these fungi are also called as “Microbial pesticides”.

Abstract

Potato (Solanum tuberosum L.), an important cash crop of India, is prone to many diseases and amongest these, late blight [Phytophthora infestans (Mont) de-Bary] is one of the highly destructive diseases and a major constraint in the profitable cultivation of potato. After realizing the significance of controlling the devastating disease for the sustainability in potato production and thus ensuring more profitability, the present investigation has been conducted with an objective to select the most effective fungicides and botanicals for the management of the disease. The present study was carried out by KVK, Saharsa during rabi season for consecutive two years (2015-16 and 2016-17) as On-Farm Trial mode at farmers fields of an adopted village of Saharsa district in Bihar. In each trial, there were four treatments including one control i.e. farmers’practice. The study concludes that the best treatment option to combat late blight disease is Seed treatment with Trichoderma viride @ 5g per kg of seed and spray of Ridomil @ 2g per litre of water starting from 30 DAP alternating with Dimethomorph 1g + ridomil 2g per litre of water at 10 days intervals. The results of economic analysis reveal that the net return as well as benefit cost ratio was also highest in Seed treatment with Trichoderma viride @ 5g per kg of seed and spray of Ridomil @ 2g per litre of water starting from 30 DAP alternating with Dimethomorph 1g + ridomil 2g per litre of water at 10 days intervals.

Abstract

Mushroom production is emerging as an added dimension in agriculture diversity; its cultivation lies on the utilization of agricultural wastes rich in proteinaceous materials. Five different substrates such as wheat straw, maize cob, paddy straw, banana pseudo stem, and lentil straw were used with suitable supplements to evaluate the average period of spawn run, pin head initiation, time taken for harvesting of flushes, fruit yield during both crop season 2014 and 2015. In 2014, highest fruit yield (380.3 g/ 500g) were recorded when wheat straw supplemented with rice bran (2% of the substrate weight), corn flour @ 25 g/kg prepared straw and gypsum @ 20 g/kg straw among various used substrates. This treatment also resulted better in respect of time taken for spawn run (9.3 days), pin head initiation (11.7 days), time taken for harvesting of flushes (I^st flush- 20.5, II^nd flush-26.7, III^rd flush- 32.9). The second best treatment was paddy straw when supplemented with rice bran (2% of the substrate weight), corn flour @ 25 g/kg prepared straw, and gypsum @ 20 g/kg. The lowest fruit yield (193.6 g/500g), time taken for spawn run (15.9 days), pin head initiation (22.4 days), time taken for harvesting of flushes (I^st flush- 27.9, II^nd flush-37.1, III^rd flush- 44.7) were recorded with banana pseudostem (chopped) when supplemented with rice bran (2 % of the substrate weight), corn flour @ 25 g/kg prepared straw and CaCO₃ gypsum @ 20 g/kg straw. The same experiment was repeated during the year crop season 2015. Surprisingly wheat straw supplemented with rice bran (2% of the substrate weight), corn flour @ 25 g/kg prepared straw and gypsum @ 20 g/kg prepared straw was found again superior over other treatments with more or less similar performance. Similarly more or less same performances were resulted in rest of the treatment too. The incidence of mould infection in mushroom was also noticed in accordance with the kinds of substrates used during both crop season 2014 and 2015. Lowest incidence of mould i.e. green mould (5.13 %), black mould (3.10 %) and blue mould (2.42%) was observed on wheat straw when supplemented with rice bran (2% of the substrate weight), corn flour @ 25 g/kg prepared straw and gypsum @ 20 g/kg straw, green mold (3.13 %), black mold (3.10 %) and blue mold (2.42 %). Similarly more or less same pattern of mould incidence was reported in the crop season 2015.

Abstract

The agricultural scientists particularly those who are in advisory service, irrespective of specialization, are often approached by the farmers with diseased samples of their crops and sought advice on plant protection strategy. The strategy includes components of control measures, doses, mode and time of application. In the present article different steps of formulation of plant disease control strategy are discussed. To further elaborate the contents, examples from integrated management strategy of mango malformation disease is cited.

Keyword: Mango Malformation

Introduction

Mango malformation, disease was first time reported from Darbhanga District of Bihar State in 1891 by G. Watt. The disease is currently prevalent in almost all the mango growing country and is thus designated as the plant disease of International importance and previously considered to be a serious problem in the northern parts of the country, has now started to appear in central India too in severe form. It is well known, the mango malformation is the most serious disease, rendering mango cultivation non-lucrative in subcontinent and other regions of the world (Ahmad et al., 2002; Ploetz, 2001). Mango malformation caused by fungus Fusarium moniliforme var. Subglutinans, are proved (Freeman 1999; Zheng and Ploetz, 2002). Recently, these isolates were described as members of a new species, F. mangiferae Britz, Wingfield and Marasas sp nov. (Britz et al., 2002). The casual organism of mango malformation grows well at lower temperature and higher relative humidity. The disease is quite common in both grafted and the seedling plants (Kumar and Chakrabarti, 1997). The symptoms manifest as bunchy top (BT) in 4-5 month old seedlings in the nursery, vegetative malformation (VM) of twigs arising from axillary buds and malformation of inflorescence (MF). Normally floral malformation is more prevalent in bearing trees than vegetative malformation. Flowers in the malformed panicle are much enlarged and crowded with hypertrophied axes of the panicle. Malformed inflorescences/ panicle of different varieties were variable in respect of their shapes, sizes, growth and compactness, depending upon the cellular structure and physiology under environmental factors (Chakrabarti et al., 1990). Generally malformed inflorescences produce no fruit, or abort already stages (Schloser, 1971) and is directly responsible for reduction in yield (Kumar et al., 1993 and Singh et al., 1998). In the present study the incidence of Floral malformation on a large number of mango cultivars was recorded over a period of 5 years with a view to identify the entries of future resistant breeding programme.

Abstract

The present investigation was carried out in okra consisting of 11 diverse genotypes (8 lines & 3 testers) and their 24 F1 hybrids. Phenotypic and genotypic correlation indicated positive association of average fruit length with average fruit weight while negative association between days taken to first flower with number of fruits per plant. The maximum positive direct effect on yield was due to number of fruits per plant followed by average fruit weight. The F₁ hybrid VRO-6 × Parbhani Kranti exhibits highest negative heterobeltiosis for days taken to 1^st flowering while D-1-87-5 × Pusa A-4 and Arka Abhay × Parbhani Kranti are earliest heterobeltiotic hybrids for days taken to 1st harvest. The hybrid Arka Abhay × Pusa A-4 represents highest heterobeltiosis for average fruit length, number of fruits/ plant and average yield. Among all parents, only Akra Abhay exhibits better GCA effect. The highest SCA effect for average yield observed in VRO-6 × Arka Anamika, while VRO-6 × Pusa A-4 for per cent infestation of YVMV. The cross Arka Abhay × Pusa A-4 incurred the highest net returns/ ha and B: C ratio (3.21) followed by that of D-1-87-5 × Parbhani Kranti and the parent Hisar Unnat gave the higher net returns/ ha with B: C ratio of 2.02 among the parents. These elite hybrids may be tested for yield and other quality traits under different agro-climatic conditions for commercial exploitation of hybrid vigour.

Abstract

Spot blotch of wheat is devastating disease as one of the yield limiting factor especially in north eastern plains zone and also in north western plains zone of India. Chemical fungicides play an important role in controlling the spot blotch in India due to non availability of spot blotch resistant commercial varieties. To identify the suitable chemicals method and time of application, various fungicides thiram 37.5% WP, carboxin 37.5% WP +, propiconazole 25% EC, tebuconazole 25% EC and mancozeb 75% WP) were evaluated either as seed treatment or foliar sprays or in combination under artificial epiphytotic in vivo conditions for two crop seasons. Seed treatment (ST) by thiram + carboxin @ 2.5 g/kg of seed along with Foliar spray (FS) of propiconazole @ 0.1% at boot leaf stage on Flag-1 leaf followed by second spray at 20 days interval (two sprays) resulted in best management of spot blotch in severe conditions. It was found that the fungicide treatment not only reduced the spot blotch disease but also increases the grain yield.

Abstract

A field experiment was carried out during rabi 2012-13 at Bihar Agricultural College farm, Sabour, Bhagalpur to assess the effect of different herbicides on microbial population dynamics of soil in winter maize (Zea mays L.). The experiment comprised of ten weed management treatments involving various herbicides (acetochlor @ 1.125, 2.25 and 4.5 kg a.i. ha^-1, atrazine @ 1.0 kg a.i. ha^-1, pendimethalin @ 0.5 kg a.i. ha^-1 + atrazine @ 0.5 kg a.i. ha^-1, alachlor @ 0.5 kg a.i. ha^-1 as pre-emergence and 2,4-Diethyl ester @ 0.6 kg a.i. ha^-1and topramezone @ 0.04 kg a.i. ha^-1+ atrazine @ 0.25 kg a.i. ha^-1 as post-emergence including weed free and weedy check laid in randomized block design with three replications. Results indicated that all the herbicides’ treatment showed significant adverse effect on microbial population in rhizospheric soil of winter maize up to 45 days after sowing. Further, there was reduction in soil microbial population occurred in all the herbicides treated plots up to said time period as compared with weed free and weedy check. Initially the soil microbial population in weed free plot was much higher than that of weedy check, however, it was found in reverse trends in rest of the time intervals as well as harvest. Thereafter, soil microbial population was gradually increasing at different time intervals and at harvest, their population increased considerably in herbicidal treated plots though the weed free and weedy check showed higher soil microbial population than other herbicide treated plots.

Abstract

Lentil (Lens culinaris Medik) is an important rabi leguminous annual herb with rich source of protein and other balanced nutrition. Lentil is susceptible to many insect-pests and diseases which causes significant yield loss which may range from 50 to 100% leading to one of the most important biotic constraints in productivity. The major fungal diseases in lentil are Fusarium wilt, Ascochyta blight and rust which causes severe losses in lentil production. Other minor diseases like botrytis grey mould, anthracnose, alternaria, stem rot, collar rot and some viral diseases also infect the lentil field under condusive environment conditions. Coming to major insect pests of lentil viz. helicoverpa, aphids, leaf weevils and lygus bugs with several noteworthy minor field pests such as thrips and cutworm with few storage pests that infest the crop. With an integrated blueprint of disease management, one can minimize risk of yield loss by following modified package and practices with need based use of pesticides, thus maximizing profit in lentil production.

Abstract

Mungbean [Vigna radiata (L.) Wilczek] is an important pulse crop grown in India. It is a short duration annual with yellow flowers and fuzzy brown pods. Majority of the mungbean grown in India is under rainfed conditions and the productivity of mungbean is not upto the mark to meet the daily dietary requirement of a populous country like India. The biotic and abiotic stresses in mungbean, majorly MYMV, powdery mildew, drought, salinity, water logging etc leaves a severe yield loss in mungbean crop. Among them, MYMV and powdery mildew diseases show yield losses accounting upto 85% and 40%, respectively in severe conditions. The present review paper is based on the study of both biotic and abiotic stresses and the recent advances in breeding for resistance to stresses. A detail explanation of major diseases and insect pests affecting mungbean are discussed along with the information of genomic resources and research done in mungbean in recent years. Recently SSR’s have been using widely for all molecular level research work like genetic diversity, constructing linkage maps for identifying QTL’s, marker assisted breeding etc. in mungbean. The resistant varieties developed against these biotic and abiotic stresses are listed which are very important for attaining a prominent yield in mungbean.

Abstract

The major losses of banana at present by Panama wilt disease pathogen of banana and its management is very typical because it is caused by a soil-borne pathogen (Fusarium oxysporum f.sp. c u b e n s e ). The micronutrients are used to manage the disease as well as it provides the healthiness of plant and increases the fertility of the soil that’s why the plant is free from the disease or less infected. An experiment of micronutrients was conducted in the laboratory of plant pathology BAU, Sabour for Invitro evaluation of the micronutrients viz. Zinc Sulfate, Ferrous Sulfate, Ammonium Sulfate, Copper Sulfate, Calcium Nitrate, Borax and Potassium Chloride against Fusarium oxysporum f.sp. cubense causing Panama wilt of banana. The results of the experiment revealed that among the micronutrients used, Borax @ 500 and 750 ppm completely inhibited the mycelial growth of FOC followed by Zinc Sulfate (45.7, 42.6, 39.2 and 18.5 mm at 100, 250, 500 and 750 ppm respectively). The incorporation of borax in the solid medium at higher concentrations viz., 500 and 750 ppm is very effective for the management of Panama wilt disease of banana. This approach leads to maintain our ecosystem sustainability for a longer time and manage the disease.

Abstract

Chickpea is one of the major grain legume crops of the tropics and subtropics. It is the second most important pulse crop after pigeon pea in India and is commonly known as gram or Bengal gram is a versatile crop among the grain legumes with a total production of 11.6 (Mt) from 13.2m ha. However, it ranks fifth in the productivity after Fababean, pea, lupin, and lentil and ranks first among the pulses both in acreage and production. Whereas, area under pigeon pea 3.90 mha with a total production and productivity of 2.89 mt and 741 kg/ha, respectively and is commonly known as red gram or Arhar is a very old crop of India. The chromosome number of pigeon pea is 2n = 22 chromosomes. They are the rich source of protein and supplies a major share of the protein requirement of the vegetarian population of the country. Chickpea accounts for 70.7% of the world Chickpea production n India followed by Australia, Pakistan, Turkey, Myanmar, Ethiopia, Iran USA, Canada and Mexico. Both crops are used as an important source of protein in human nutrition and cattle feed. Chickpea is considered to have medicinal effects and it is used for blood purification. Chickpea contains 21% protein, 61.5% carbohydrate, 4.5% fat. Its seed is also rich in protein, starch, fiber, calcium, iron and niacin, malic and oxalic acid, which makes it one of the best nutritionally balanced pulses for human consumption. However, like any other pulses, the chickpea seed also contains antinutrional factors, which can be reduced or eliminated by cooking. Both crops enrich the soil fertility by fixing atmospheric nitrogen in the root nodules and improve the soil structure. It is also to improve soil fertility by biological nitrogen fixation which helps reduce the input cost for the existing crop. These crops usually receive few inputs other than labor, insecticides and seed. Chickpea is a crop of both tropical and temperate regions. Low yield of chickpea attributed to its susceptibility to several fungal, bacterial and viral diseases. In general, estimates of yield losses by individual insects and diseases range from 5 to 10% in temperate regions and 50 to 100% in tropical regions. Whereas, Pigeon pea can be attacked by more than 100 pathogens. These include fungi, bacteria, viruses, nematodes, and mycoplasma like organisms. Fortunately, only a few of them cause economic losses and the distribution of the most important diseases is geographically restricted. The diseases of considerable economic importance at present are Fusarium wilt, sterility mosaic, phytopthera blight and alternaria blight.

Abstract

Fusarium wilt is major disease in Central India, the major lentil growing region of country. The disease is soil borne causing heavy loss, development of wilt resistant varieties is most effective means of controlling this disease. Highly resistant sources of wilt in lentil have not been reported from the Indian lentil breeding programme. One hundred thirty lentil genotypes including one susceptible check were evaluated in control conditions of green house against wilt, based on controlled condition screening- LL 1370, IPL 333, PL4, RLG 195, RVL 14-4, L 4737, RKL 607-1, WBL 77, LL 1320, LL 1397, VL 525, LL1379, VL526, RVL14-5, L4717, VL126, L4076, RLG191 and VL150 have been identified as highly resistant genotypes. These can be used in hybridization programme for wilt resistance breeding and studying the inheritance of wilt resistance in lentil.

Abstract

Lentil is an important pulse crop and play very important role in the supply of the protein to under nourished vegetarian population of the country. Fusarium wilt of lentil caused by Fusarium oxysporum f. sp. lentis is one of the wide spread and destructive diseases in India under rice-based cropping system. Use of synthetic fungicides has led to the emergence of several problems like environment pollution, residual effect in grains and killing of non-target organism(s). Hence, for minimizing the losses caused by Fusarium wilt, four plant extracts such as Garlic, Neem, Eucalyptus, Tulsi and three antagonistic bio-agents viz. Trichoderma viride, Trichoderma harzianum and Pseudomonas fluorescence were evaluated in vitro. The results clearly showed that all plant extracts were more or less inhibitory to mycelial growth of Fusarium oxysporum f. sp. lentis. The effectiveness of extracts increased with increase of their concentration and maximum growth inhibition was obtained in Eucalyptus followed by Garlic and Tulsi. While minimum per cent inhibition was recorded in Neem. Among bio-agents, T. viride showed maximum growth inhibition of Fusarium oxysporum f. sp. lentis followed by T. harzianum at both 4 and 7 days incubation period.

Abstract

Pigeon pea is an important pulse crop of India and its production is greatly affected by wilt disease. Pigeon pea wilt is a very destructive soil borne disease caused by Fusarium udum. In present research work plant extracts (Garlic, Neem, Tulsi), bio-agents (Trichoderma viride and Pseudomonas fluorescens) and fungicides (Vitavax, Bavistin and Thiram + Carbendazim) were tested against Fusarium udum. Among them fungicides were found most effective in inhibiting mycelial growth of Fusarium udum recorded 100% inhibition followed by garlic bulb extract (52.12 to 56.33%). Among bioagents, Pseudomonas fluorescens inhibit the growth up to (46.00 to 51.79%) followed by T. viride (44.32 to 48.79%) amongst all treatments except untreated.

Keywords: Pigeon pea, Fusarium udum, bio-agent, fungicide

Pigeon pea (Cajanus cajan L. Millsp.) is one of the major pulse crops grown in the tropics and subtropics belonging to family Leguminaceae. It has wide adaptability and low input requirements mostly grown in kharif season.

Abstract

Pigeonpea (Cajanus cajan) is an important pulse crop of India and its production is greatly affected by wilt disease. Pigeonpea wilt is a very destructive soil borne disease caused by Fusarium udum. In the experiments, ten treatments were applied viz.; Neem leaf extracts (T₁), Garlic extracts (T₂), Tulsi leaf extracts (T₃), bio-agents Trichoderma viride (T₄), Pseudomonas fluorescens (T₅), Polymer coating (polycot) @ 4 ml diluted in 5 ml of water per kg of seed (T₆), Polymer coaating + Trichoderma viride @ 6.0 g per kg seed (T₇), Polymer + Pseudomonas fluorescens @ 6.0 g per kg seed (T₈), Polymer coating (polycot) 4 ml diluted in 5 ml of water per kg seed + Vitavax power @ 3g per kg seed (T₉) and T₁₀ served as untreated. The observations were recorded six times at 10 days intervals and started from 40 days. These polymer alone @ 4.0ml per kg and in combination with bioagents and fungicides were applied to seed were tested against Fusarium udum. The highest per cent disease control and per cent wilt disease incidence were recorded 96.09 to 64.09 and 2.74 to 33.32, respectively in T₉ (Polymer + Vitavax power @ 3g per kg seed) at 10 days intervals followed by T₈ (Polymer + P. fluorescens) gave 94.46 to 53.06 per cent inhibition, T₇ (92.39 to 52.74) and T₆ (90.97 to 49.73) however, the lowest per cent disease control was noticed as 76.19 to 40.13 per cent in T₁ (Neem leaves extract) amongst all treatments except untreated.

The concept of gene has undergone several refinements since its discovery in 1866 by Mendel. Most genes encode one polypeptide and can be operationally defined by the complementation test. The complementation or cis-trans test provides an operational definition of the gene. It is used to determine whether mutations are in the same gene or in different genes. Intragenic complementation may occur when a protein is a multimer containing at least two copies of one gene product. The concept of gene has evolved from a bead on a string, not divisible by recombination or mutation to a sequence of nucleotide pairs in DNA encoding one polypeptide chain. The unit of genetic material not divisible by recombination or mutation is the single nucleotide pair. Historically several experimental developments have occurred to facilitate understanding of the structure and function of gene and these are briefly described in a chronological order as follows to have better understanding of the gene concept as it evolved. However, before discussing evidences supporting various gene concepts, it is better to summarize important stages in the evolution of the gene concept.

Regulator Genes

Bacterial cells are capable of producing many kinds of enzymes in relatively large quantities but the kinds and amount of enzymes actually produced at any given time are usually those which are required at that particular time in response to the prevailing environment. Inducible enzymes are ordinarily absent or present in very small quantities until a specific compound appears. This compound is called as inducer. A typical example of this is production of b galactosidase (an enzyme that splits lactose into galactose and glucose, Fig. 44.1) once the lactose is introduced into the medium on which E. coli cells are grown. Here lactose acts as inducer for the enzyme b galactosidase.

Biochemical reactions are mediated by enzymes and all enzymes are proteins. Proteins are polymers of subunits (monomers) called amino acids which are often termed as ‘residues’. Each amino acid has an amino group (NH ) at one end and a carboxyl group (COOH) at the opposite end. 20 different kinds of amino acids occur naturally in proteins. Each enzyme consists of a certain number of amino acids in a precisely ordered sequence. The blueprint for making proteins is coded in the nucleotide sequence of DNA. The number of nucleotides which code for an amino acid is termed a codon. There are 20 common amino acids but only four different nucleotides. Obviously a singlet code (one nucleotide coding for one amino acid) could code only for four amino acids. A coding sequence of two nucleotides for one amino acid or a doublet code would produce 4₂ = 16 possible coding combinations or codons and this arrangement is not sufficient to take care of 20 amino acids. A codon size of three nucleotides for one amino acid or triplet code seems more likely as it produces 4₃ = 64 possible codons as illustrated in Table 43.1 (Strickberger, 1968).

In transcription, it has become clear as to how there is the transfer of genetic information stored in the sequence of nucleotides-pairs in DNA to the sequence of nucleotides in m-RNA molecules which carry that particular information from the nucleus to the sites of protein synthesis (ribosomes) in the cytoplasm. Further, there is the requirement of the process by which genetic information stored in the sequences of nucleotides in m-RNAs is used to specify the sequences of amino acids in the polypeptide gene products. This process known as translation takes place in the cytoplasm on complex workbenches called as ribosomes. With exception of water, proteins are by far the most prevalent component of living organisms in terms of total mass.

Proteins (= enzymes = genes) are composed of polypeptides and every polypeptide is encoded by a gene. Each polypeptide has a long sequence of amino acids linked together by covalent bonds. There are 20 different amino acids (glycine, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan, cysteine, serine, threonine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine, and histidine) present in most proteins. All amino acids except proline contain a free amino group (NH₂) and a free carboxyl group (COOH) as illustrated below:

According to the central dogma of molecular biology, genetic information flows (i) from DNA to DNA during its transmission from generation to generation and (ii) from DNA to protein during its phenotypic expression in an organism (Fig. 41.1).

Watson and Crick based on their double helical DNA model, offered to explain DNA replication by a method where each single strand of DNA is template or mold for its complement and a new helix has one old strand and one newly synthesized strand. This type of DNA replication is called as semi-conservative as in this method one old strand is retained and one new complementary strand is synthesized. In contrast to semi-conservative mode of DNA replication, two more modes of DNA replication were proposed. One is conservative type where two new strands are synthesized in form of a double helix while the old double helix remains unchanged. The third type of DNA replication proposed has been dispersive mode of DNA replication which amounts to breaking down of double helical strands into small pieces along the length, their replication and random assemblage with newly synthesized pieces to form a patchwork single string of dispersed old and new pieces. These three modes of replication of DNA molecule are shown in Fig. 40.1.

The search for the chemical basis of genetic material began about one and half century ago. Soon after discovery of Mendelian principles of segregation and independent assortment in 1865, Friedrich Miescher (1844-1895) published a method for separating cell nuclei from the cytoplasm and from these cell nuclei, he extracted an acid material termed as “nuclein” which had unusually large amount of phosphorous. Miescher concluded that this substance was unique and was just not comparable with any other group known at that point of time. Later on nuclein, now named as “nucleic acid” was found to be associated with various proteins and the combined entities were termed as “nucleoproteins”.

In 1868, Johann Friedrich Miescher, a young Swish medical student got excited with an acidic substance that he isolated from pus cells obtained from bandages used to dress human wounds. To obtain large quantities of pus for his experiment, Miescher began purchasing used bandages from surgical clinics in Tuebingen, Germany. He separated pus cells from bandages and treated the cells with pepsin, a proteolytic enzyme that he isolated from stomach of pigs. After pepsin treatment, he recovered an acidic substance that was named as “nuclein” by him. This nuclein was unusual in the sense that it was rich in nitrogen and phosphorous, two elements known at that time to coexist only in certain types of fat. Miescher’s paper on this investigation was published in 1871. At that time, importance of Miescher’s nuclein was not fully appreciated. Even the existence of polynucleotide chains, the key component of the acidic material in Miescher’s nuclein was not documented until 1940s. The role of nucleic acid in storing and transmitting genetic information remained un-established until 1944 and the double-helix structure of DNA was not discovered until 1953 by James Watson and Francis Crick. Even in 1953, many geneticists remained skeptical to accept the role of nucleic acid as the genetic information carrier rather than proteins as nucleic acids exhibited less structural variability than the proteins (Snustad et al., 1997).

So far, we have seen the examples in which mode of inheritance of a particular character is tied to the behaviour of the nucleus or more specifically to the chromosomes. In other words, transmission of a character and its appearance in the individuals can be predicted from the knowledge of chromosome segregation and independent assortment. This analogy gets further strengthened on the ground that the DNA has been considered as the basic genetic material and practically all the cellular DNA is localized in the chromosomes. It has also been seen that DNA does not produce an organism alone but obviously depends upon an already existing medium in order to function. Thus, genotypic effects may considerably get modified by the environment. In the cell itself, one important source of environmental effect is the cytoplasm immediately surrounding the nucleus. The components of the cytoplasm may show wide variation between the individuals and therefore it cannot be ruled out that a genotype in one cytoplasm may function somewhat differently than the same genotype placed directly into another cytoplasm. It is well known fact that the cytoplasmic contribution of one of the parents in form of an egg is considerably larger than the cytoplasmic contribution of the other parent in form of sperm. For example, cross-fertilization between two strains capable of producing both sperm and eggs, leads to contribution of ‘A’ genotype and ‘a’ egg cytoplasm in one case and of ‘B’ genotype and ‘b’ egg cytoplasm in the other case. Thus, the cross-fertilized diploid zygotes have two possible genotypic-cytoplasmic combinations, namely, AB-a, and AB-b depending upon which strain provides the egg. If the egg is contributed by the A parent, then the genotype of the progeny shall be AB-a, and if the egg is contributed by the B parent, then the genotype of the progeny shall be AB-b. If each such cytoplasm produces a unique phenotype, then AB-b would have the same b phenotype as BB-b individuals. Such effects produced by the egg cytoplasm are termed as maternal effects.

Translocations

Chromosomes occasionally undergo spontaneous rupture or can be induced to rupture in high frequency by ionizing radiation. The broken ends of such chromosomes behave as though they were ‘sticky’ and may rejoin into non- homologous combinations giving rise to translocations. The most common type of translocation is reciprocal translocation which involves the exchange of segments between two non-homologous chromosomes. Translocations change only the arrangement of genes in the chromosomes, not the quality or quantity of genes. For this reason, they are sometimes referred to as chromosomal rearrangements. Individuals carrying such rearrangements should be phenotypically entirely normal unless the relations of a gene or genes to adjacent genes affect the phenotypic expression (position effect). The first translocation was discovered by Bridges in 1923 in D. melanogaster.

The genetic techniques for detecting and studying translocations will be more easily understood when the cytological phenomena produced by translocations are known. Suppose two chromosomes having respectively the genes ABCDEF and GHIJKL exchange sections and give rise to translocation chromosomes ABCJKL and GHIDEF. The individual thus formed receives from one of the parents the normal and from the other parent the translocation chromosome. Such an individual is a translocation heterozygote (Fig. 36.1).

When there is indication of linkage from F or testcross data through significant deviation from 9: 3: 3: 1 and 1: 1: 1: 1 ratio, it is implied that independent assortment is not taking place and this is taken as an indication of linkage. The inverse of linkage strength is the recombination value and is often symbolized by ‘p’ whose maximum value is 50% because out of four strands of a tetrad, two are always parental types and two are recombinant types when all the cells show crossing over.

Linkage

When two or more genes are located on the same chromosome, they are said to be linked. Genes on different chromosomes are distributed to the gametes following the law of independent assortment. However, genes located on the same chromosome, tend to stay together during gamete formation. For example, results of test-crossing dihybrid individuals will produce different results depending upon whether the genes are linked or are located on the same chromosome. This is illustrated as follows:

In most of higher organisms including animals, the aspect of sexes has been normally confined to males and females which exist separately or even in the same individual. An individual possessing both male and female reproductive organs is usually referred to as hermaphrodite. In plants, where staminate (male) and pistillate (female) flowers occur on the same plant, the term used is monoecious. The most common plants under monoecious form are cucurbits. However, majority of the flowering plants have male and female parts within the same flower and this situation is referred to as hermaphrodite. There are relatively less numbers of plants having male and female flowers located on different plants and this is termed as dioecious condition. Among the common cultivated crops known to be dioecious are asparagus, date palm, hemp, hops and spinach.

Phenotype is a result of gene products brought to expression in a given environment. Genes specify the structure of proteins. All known enzymes are proteins. Enzymes perform catalytic function, causing the splitting or union of various molecules. All of the chemical reactions which occur in the cell constitute the subject of intermediary metabolism. These reactions occur as stepwise conversions of one substance into another, each group being mediated by a specific enzyme. All of the steps which transform a precursor substance to its end product constitute a biosynthetic pathway. Several genes are usually required to specify the enzymes involved in even the simplest pathways. Before gene-enzyme relationship could be postulated, soon after rediscovery of Mendelian principles of inheritance, it was discovered that genes were not merely separate elements producing distinct individual effects, but they could interact with each other to produce completely novel phenotypes. The simplest example of gene interaction could be the fact that allele A could be characterized by phenotype A, and the allele B by the phenotype B and an organism having both alleles A and B at the same time might show phenotype C. The occurrence of such interactions means that the phenotype we observe is not present in the genes themselves but arises from a complicated developmental process resulting from a biosynthetic pathway as illustrated below (Stansfield, 1969)

Historically the first and still the most conclusive evidence for the existence of genes comes from the phenomenon of segregation of traits observed in the offspring of hybrids between individuals or strains that differ in some recognizable respect. The principle of segregation was formulated by Gregor Johann Mendel in 1866 under such peculiar circumstances that the scientific world failed to recognize it until after a lapse of 34 years. In the first place Gregor Mendel was not primarily a biologist but a Monk in the Augustinian monastery at Bruenn, Austria (now Brno, Moravia in Czech Republic). During his childhood, Mendel worked as a gardener and studied beekeeping. He became part time monk because it enabled him to obtain an education without having to pay for it himself. From 1840 to 1843 he studied Practical and Theoretical Philosophy and Physics at the Philosophical Institute of the University of Olomouc. Upon recommendation of his teacher, Mendel entered the Augustinian St. Thomas Abbey in Brno and began his training as a Priest. Born Johann Mendel, he took the name Gregor upon entering religious life. Mendel worked also as a substitute High School teacher. In 1850, he failed the oral part of his examination and could not become a certified High School Teacher. In 1851, he was sent to the University of Vienna to study further so that he could get formal education. Mendel returned to his Abbey in 1853 as a Physics teacher. In 1856, he took the examination to become a certified teacher but again failed the oral part.

Chi-Square

The Chi-square test (c₂) proposed by Karl Pearson in 1899 is used to measure goodness of fit of observed and theoretical or expected data and is a continuous distribution.

Formula for Chi-Square

1. General Formula

The general formula for Chi-square is as follows

Cell

Cells (small, membrane-bounded structures filled with a variety of chemicals in an aqueous solution that interact to give life) are the basic unit of life. Cells are capable of acquisition and use of energy, have the capacity to reproduce and respond to the environment and are able to carry out a variety of chemical reactions. They maintain a relatively constant internal environment despite changes in the external environment. There are two types of cells, namely, prokaryotes and eukaryotes.

The prokaryotic (pro = before, karyote = nucleus) cells are the simple form of cell types which include the various types of cells of bacteria and their relatives, all belonging to kingdom Monera. They possess a cell wall constructed of peptidoglycan, a substance unique to prokaryotes. Beneath the cell wall, there is a plasma membrane that encloses a cytoplasm containing DNA, RNA, proteins and various small molecules.

Genetics and Gene

Genetics is defined as the branch of biology dealing with heredity and variation. The hereditary units which are transmitted from one generation to the next are called genes. Genes have been defined in various ways. These are units of inheritance located in linear order on chromosomes. The genes are considered as hereditary determinants of a specific biological function located in a fixed place on the chromosome. Chemically, gene is a segment of DNA (deoxyribonucleic acid) encoding one polypeptide and defined operationally by the cis-trans or complementation test. Gene is the functional unit of inheritance (a sequence of DNA) which is located on a certain position (locus, plural loci) on a chromosome. The locus is sometimes used interchangeably for gene. DNA, in conjunction with a protein matrix, forms nucleoprotein and becomes organized into structures with distinctive staining properties which are known as chromosomes. The chromosomes are located in the nucleus of the cell. Since genes are located on the chromosomes, in many ways behavior of genes is parallel to the behavior of the chromosomes. Two or more alternative forms of a gene are called as alleles. All genes on a chromosome are said to be linked and they all belong to the same linkage group.

Varietal release and notification comes under the purview of the Seeds Act which is intended to provide for regulating the quality of certain notified seeds for sale and for marketing and for the matters connected therewith. Released and notified varieties are offered for seed certification by the seed certification agencies which ensure that the varieties under certification meet certain prescribed field and seed standards failing which they stand to be rejected during the process of seed certification.

Purpose of Release

The purpose of release of cultivars is to introduce the newly evolved varieties to the public for general cultivation in the region in which it is suitable. It enables the farmers to choose cultivars for cultivation in a region. In other words, release of a cultivar is in the nature of a recommendation to the farmers for its adoption. Therefore, notification of a variety is linked to the release of a variety, though the release process itself does not have statutory cover.

Plant breeding activities basically involve development of new and superior crop varieties to replace the old ones and this naturally needs comparison of several lines/cultivars for agronomically important traits which are often quantitative in nature. Therefore, lots of environmental influences are there on the mean performance and mere numerical comparison of mean data is not sufficient to draw valid conclusions regarding the superiority of one variety over the other. Great advances have been made during the last about 100 years in the area of experimental designs and analytical procedures to facilitate such comparisons. Most commonly used designs by the plant breeders are included here for better understanding by the students and the breeders and to handle the experimental data on their own when confronted with such issues. Before applying statistical procedures on the experimental designs, it must be well understood that proper use of field plot technique is of great value to have meaningful data for further statistical analysis and use.

Statistics

Statistics may be defined as the mathematical analysis, based on the theory of probability, of experimental data in an attempt to summarize or to describe them so that conclusions can be drawn with respect to the phenomena that supply the data. It is the branch of mathematics concerned with collection, classification, analysis, and interpretation of numerical facts, for drawing inferences on the basis of their quantifiable likelihood (probability). Statistics can interpret aggregates of data too large to be intelligible by ordinary observation because such data (unlike individual quantities) tend to behave in regular, predictable manner. It is subdivided into descriptive statistics and inferential statistics. Statistics is also defined as a discipline that allows researchers to evaluate conclusions derived from sample data. Plural of statistic is also statistics. In practice, statistics refers to a scientific approach used to:

There have been lots of international efforts to establish crop based research centres primarily in the developing/under-developed countries to carry out concerted breeding and plant genetic resources management activities pooling germplasm from all over the world and also engaging scientists from different countries to do breeding work on the certain mandate crops of the particular region using all kinds of germplasm and technologies with strong financial support from a central agency known as Consultative Group on International Agricultural Research (CGIAR) for doing the stipulated work as per programme and deliver the products which can be used by different national plant breeding programmes either to use them directly as cultivars or to use them in their breeding programmes in various ways including parents in the crossing programme.

CGIaR (Consultative Group on International agricultural Research)

CGIAR System Organization headquartered in Montpellier, France is a global agricultural research partnership for a food secure future that was founded in the 1970s in response to the Green Revolution. There are presently 15 CGIAR agricultural research centres employing more than 8,500 researchers and support staff worldwide, with an annual budget of US$ 800 million.

CGIAR is a global research partnership for a food-secure future. CGIAR science is dedicated to reducing poverty, enhancing food and nutrition security, and improving natural resources and ecosystem services. Its research is carried out by 15 CGIAR Research Centers in close collaboration with hundreds of partners, including national and regional research institutes, civil society organizations, academia, development organizations and the private sector. The CGIAR Fund was established as a Financial Intermediary Fund (FIF) in 2010. It was subsequently closed and replaced by a new FIF in 2017, following the CGIAR System governance reorganization.

Biotech crops or genetically modified (GM) crops or the transgenic crops are those crop plants into which certain genes have been introduced from unrelated sources beyond the boundary of sexual compatibility using techniques which bypass the normal sexual cycle. The transgenic production technology uses recombinant DNA technology where a recombinant DNA molecule is a vector into which the desired DNA fragment has been inserted to enable its cloning in an appropriate host. The recombinant DNA technology has opened up the flood gate of plant biotechnology to produce plants which are equipped with novel genes to produce products never heard of that a plant will do. Now transgenics in several crops have been commercialized globally. The importance and potential of GM crops can be realized from the fact that acreage under such crops at global level has reached to 189.8 million ha covering 24 countries in 2017. One can have a feel of this huge acreage under commercialized GM crops globally if it is compared with about total cultivated area of 143 million ha in India.

Over the last 30 years field of agricultural biotechnology has expanded rapidly due to greater understanding of DNA as the chemical double helix code from which genes are made. Genetic engineering is one of the modern biotechnology tools that are based on recombinant DNA technology. The term genetic engineering, often interchanged with terms such as gene technology, genetic modification, or gene manipulation, is used to describe the process by which the genetic makeup of an organism can be altered using “recombinant DNA technology.” This involves the use of laboratory tools and specific enzymes to cut out, insert, and alter pieces of DNA that contain one or more genes of interest. The ability to manipulate individual genes and to transfer genes between species that would not readily cross is what distinguishes genetic engineering from traditional plant breeding and opens up a huge opportunity to create crop plants equipped with desirable genes from the sources which cannot be sexually crossed. With conventional plant breeding, there is little or no guarantee of obtaining any particular gene combination from the millions of crosses generated. Undesirable genes can be transferred along with desirable genes or while one desirable gene is gained, another is lost because the genes of both parents are mixed together and re-assorted more or less randomly in the offspring. These are the major limitations of conventional crop breeding.