eChapters

Diagnosis of plant diseases require some knowledge of light microscopy to examine infected tissues for the presence of plant pathogens. Most of the pathogens and the diseases caused by them can be identified quickly by using different methods such as section or cutting of infected tissue, squashes of scrapping of plant surface. Homecare, the detail study enquires ultrastructural examination of the pathogen.. Electron microscope rely on a beam of electrons produced from a hot filament (e.g. tungsten) and its power of resolution is much greater than light microscope. Electron microscope are of two type

For growing fungi, a wide range of media are used. Media affect colony morphology and color. For utilizing different carbon sources some fungi lack the necessary enzymes. For growth and reproduction all fungi require several specific elements. The requirements for growth are generally less specific than for sporulation, so it is often necessary to try several types of media when attempting to identify a fungus in culture. Sporulating cultures should be used because repeated transfers of a pathogen on artificial media usually result in loss of sporulation or pathogenicity or both. Transfer should be made using spores and cultured alternately on nutrient rich and poor culture media. Most fungi thrive on potato dextrose agar (PDA), but in some fungi excessive mycelial growth is obtained at the expense of sporulation. Similarly, wood-inhabiting fungi and dematiaceous (dark pigmented) fungi often sporulate better on Corn meal agar (CMA) or Oat Agar, both of which have less easily digestible carbohydrate than PDA. Cellulose-destroying fungi grow on a weak medium such as Water Agar (WA) or Potato Carrot Agar (PCA). By adding pieces of tissue, such as filter paper, wheat straw, rice, grains, leaves or dung, often produces good sporulation dependent on the organism grown. Some special methods of inducing sporulation in fungi.

In the classification given below only those groups have been given emphasis which contain plant pathogenic fungi or which are important from taxonomic point of view. The description includes only critical features.

There are great variations in the life cycles of fungi. Raper (1954, 1966b) recognizes seven basic types.

5.1 Asexual cycle

In which sexual reproduction apparently lacks and diploidy is restricted to somatic diploids. The entire groups known as Fungi Imperfecti and numerous other species which clearly belong to various groups of the perfect fungi such as Penicillium notatum have this life cycle. Certain benefits of sexuality are provided here in many of the sterile forms by para sexual recombination.

Reproduction in fungi is the formation of new individuals having all the characters of species. It is of two types. Asexual, somatic, or vegetative reproduction does not involve the union of nuclei of sex cells or sex organs. Sexual reproduction is characterized by the union of two opposite nuclei. During reproduction entire thallus is converted into one or more reproductive units. It is called holocarpic fungi (holo = whole, korpos = fungi) ex. Plasmodiophora and Synchytrium. In Plasmodiophora vegetative body is plasmodium, which is transformed into reproductive structure. In some fungi only a part of thallus is transformed into one or more reproductive structures. (eu = true, good, carpus = fruit). These fungi are known as eucarpic. Both somatic and reproductive structures coexist. Ex. Rhizopus, Puccinia and Helminthosporium.

Fungi are achlorophyllus so they have to depend on other organisms hence heterotrophic nutrition. Depending on the nutrition, fungi are classified as Saprophytes, Parasites and Symbionts.

3.1 Saprophyte (sapros = rotten; phyton = plant)

Fungi which get their nutrition only from dead organic are called saprophytes ex: Saprolegnia, Rhizopus, Mucor, Alternaria etc.

3.1.1 Facultative saprophyte (Facultas = ability)

Ability of a fungus to become saprophyte. It is a parasite ex. Ustilago

3.1.2 Obligate saprophyte (obligate = to bind itself)

When they strictly get their food only from dead organic matter and can never become parasites are called obligate saprophyte ex. Mucor.

According to G.C. Ainsworth (1973) the fungus is free-living, parasitic or mutualistic, saprotrophs, necrotrophs or biotrophs and devoid of chlorophyll. Cell wall composition very variable, majority contain chitin and glucans, reserve materials glycogen, oil and mannitol, characteristic disaccharide and trehalose. Some yeast-like but majority with thread-like filaments, hyphae, branched profusely to form the vegetative mycelium on which spores are produced, asexually or sexually, free on hyphae or enclosed in complex reproductive structures. Separated mainly on morphology of the latter.

2.1 Divisions: Myxomycota

These are wall-less and quite unusual organisms only included in the fungi as mostly studied by mycologists. Possess either a plasmodium, a mass of naked, multinucleate protoplasm, which feeds any ingesting particulate matter and moves by amoeboid cells. Both of slimy consistencies hence slime molds. Four classes:

2.1.1 Class: Acrasiomycetes (Cellular slime molds)

Assimilative phase, free living amoebae which aggregate to form a pseudoplasmodium before reproduction.

1.1 Introduction

Fungi are chlorophyll-less thallophytic plant. Due to absence of chlorophyll, they are heterophytes which depend on other form of life for food. They grow in various habitats and show much diversity in their structure, physiology and reproduction. They evoloved long back from the ancient time.

Their existence was recorded from Pre-cambrian period. Information from ancient literature indicates that the fungi were used as food by men. At present, the fungi are used in medicine as well as food in addition to other aspects. The fungi cause diseases in crop plants (spots, rusts, smuts etc.) and on human beings (Aspergillosis, Blastomycosis etc.).

At present biologists use the term fungus (fungus mushroom, from Greek, sponagos = sponge) to include eukaryotic, spore bearing, achlorophyllous organism that generally reproduce sexually and asexually and whose usually filamentous, branched somatic structure are typically surrounded by cell wall containing chitin or cellulose or both of these substances, togather with many other complex molecules

More than 1.5 million species (Hawksworth 2001; Kirk et al., 2001) of fungi have been recorded, but their number may be much more than the actual record. The subject which deals with fungi is known as Mycology (mykes — mushroom; logos—study) and the concerned scientist is called mycologist. The various definitions of fungi as proposed by mycologists are:

Alexopoulos (1962) defined fungi as “nucleated, spore-bearing, achlorophyllous organisms which generally reproduce sexually and asexually and whose usually filamentous, branched somatic structures are typically surrounded by cell walls containing cellulose or chitin or both”.

Alexopoulos and Mims (1979) defined fungi as eukaryotic spore bearing, achlorophyllous organisms that generally reproduce sexually and asexually, and whose usually filamentous, branched somatic structures are typically surrounded by cell walls containing chitin or cellulose, or both of these substances, together with many other complex organic molecules.

Plant nutrients are important for increased production of food for the ever-increasing world population. Of the plant nutrients, primary nutrients N, P and K are applied to ?elds each season in fairly large amounts. In addition to increasing crop yields, they also impact the environment. This is briefly discussed in this chapter; only main points are made here.

NITROGEN

Nitrogen is blamed the most for environmental degradation. It contributes to eutrophication of inland and sea-waters, enrichment of air with NH₃ and N₂O and even with global warming. However, it is almost impossible to grow more food without additional nitrogen. It is estimated that by 2050 about 250 million tonnes (MT) of fertilizer nitrogen may be needed to meet global increased food demands as against an estimated current consumption of 116 MT

The introduction of high-yielding varieties of wheat and rice, the development of intensive cereal–cereal rotations (rice–wheat, rice–rice, hybrid maize–wheat), application of high doses of N (above 100 kg N ha^–1 to each crop) without adequate P and K application, increased use of high-analysis fertilizer (urea and diammonium phosphate in place of ammonium sulphate and superphosphate) and ignoring the application of organic manures led to heavy depletion of P, K, S, Zn and other plant nutrients, which led to wide-spread de?ciencies of P, S and Zn in Indian soils. These de?ciencies of nutrients led to focus on integrated nutrient management.

Fertilizer recommendations are generally made on the basis of soil test. Recommendations of fertilizer for top-dressing are best made on tissue testing.

SOIL TEST

A number of approaches have been adopted to increase the precision of soil-test values in making fertilizer recommendations. Four most important ones are discussed.

Low-Medium-High Approach

Making fertilizer recommendations based on soil test for a nutrient involves conducting a large number of ?eld experiments on soils similar to those which are intended for making recommendations. For an individual nutrient (say P for example) experiments are conducted with graded doses of P on a crop for which the fertilizer recommendations are to be made. Thus, agronomic experiments are to be conducted for each crop on each kind of soil. The response curve can then be divided into three sections: a, low soil-test value and high response to fertilizer zone; b, medium soil-test value and medium response to fertilizer zone; and c, high soil-test values and no response to fertilizer zone. In India, this was done

Soil-rejuvenation power of legumes in crop rotations has been known in several parts of the world since ages. It is mentioned in the Indian agriculture of the Chalcolithic period (ca 1000 BC) (Raychaudhuri and Mira, 1993), in Chinese agriculture in the Han dynasty (ca 2nd century) (Hsu, 1980) and in Roman agriculture (White, 1970). Legumes can be ?tted in cropping systems in India in a number of ways, such as, a crop in a cropping system, a catch crop in an intensive cropping system, an intercrop and as a green manure. A brief discussion on these follows.

Before the discovery of chemical fertilizer in early 20^th century, organic manures and growing of legumes were the ways of replenishing soil fertility. The only other way was leaving land fallow for a few years before its re-cultivation, a practice followed in shifting cultivation (jhuming) (Garbyal, 1999). This was also true for India, where although the ?rst single superphosphate plant was set up in 1906 in Ranipet (Tamil Nadu), the real use of fertilizer started with the manufacture of urea, the ?rst plant for which was set up in 1959 at Sindri (Bihar) (Tandon, 2014). Organic manures could meet crop-nutrient needs in early days because the average grain yield of most cereals was about 1.5–2.0 t ha^–1, while the average yield of pulses was 0.5–0.75 t ha^–1. However, the crop production scenario changed rapidly after the introduction of dwarf, high-yielding varieties of wheat from Mexico, which led to the “Green Revolution” (Swaminathan, 2013). This was followed by the development of semi-dwarf, high-yielding varieties of rice and hybrids of maize, sorghum and pearl millet. These varieties/ hybrids of cereals needed 100–150 kg N ha^–1 in one growing season, and this could be met only by urea or other N fertilizers; organic manures had no chance and their use became almost nil over a period of next 25 years or so. Well-off young farmers cannot even think of making manure heaps or compost pits, the job is too hard and not a clean one. However, what is ignored is the fact, that organic manures not only supply plant nutrients but also build up soil organic matter, which improves soil physical properties, helps in storing moisture and is the elixir of life for the soil biota (Lal, 2014). A large numbers of reports are available on the effect of adding organic manures and crop residues on improving soil physical (Hati et al., 2007; Brar et al., 2015) and chemical and biological properties of soil (Prasad et al., 1999; Prasad and Mishra, 2001;Sharma et al., 2009; Prasad et al. 2014). Thanks for the organic farming (Prasad, 2005), the interest in organic manures and composts has increased in India and it is now a sought after material. However, it has not entered the main stream of crop production.

Boron (B) de?ciency has been realized as the second most important micronutrient constraint in crop production after that of Zn in the world (Ahmed et al., 2012). However, in Nepal, B de?ciency is more wide-spread than Zn de?ciency, as 80 to 90% of soil samples were de?cient in B, 20 to 50% in Zn and 10 to 15% in Mo (Anderson, 2007). In India, B de?ciency has increased from 2% of the soil samples de?cient in B in 1980s (Katyal and Vlek, 1995) to 52% soil samples in 2000s (Singh, 2012). Total B content in Indian soils varies from 7 to 630 mg kg^–1 soil (Kanwar, 1976); being the highest in arid regions of the country and the lowest in wet humid regions of the country (Behera et al., 2009; Prasad et al., 2014). Boron de?ciency is most in the heavy rainfall areas of Western Ghats of Maharashtra, Karnataka and the eastern states of Bihar, Odisha, Jharkhand and Chhattisgarh states of India (Fig.13.1). The wide-spread de?ciency of B in India can be seen from the fact that B fertilization recorded the highest bene?t : cost (B : C) ratio in rice–wheat cropping system at Palampur (Himachal Pradesh), Ludhiana (Punjab), Faizabad, Modipuram and Varanasi (Uttar Pradesh), Pantnagar (Uttarakhand) and in rice–rice cropping system at Maruteru (Andhra Pradesh), Karjat (Maharashtra) and Jorhat (Asom) (Singh and Goswami, 2014). Response of wheat to B fertilization is also reported from Bangladesh (Halder et al, 2007).

Internationally maize (Zea mays) and citrus (Citrus sp.) are reported to largely suffer from Zn de?ciency, for which these are considered as marker plants. Zinc de?ciency in maize results in pale or whitish top young leaves and the de?ciency is referred to as 'white bud' (Camberato and Maloney, 2012). In citrus, Zn de?ciency results in ‘rosette’ (cluster of stiff leaves at the end of young shoots) formation (Brennan, 1993). Zinc de?ciency in India was ?rst reported in rice by Nene (1966) at the Govind Ballabh Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, and given the name ‘khaira’ disease due to dark-brown-coloured spots resembling kattha (Acacia catechu extract used in paan – the betel leaves) on the leaves of zinc de?cient plants. Soon All India Coordinated Project on Micronutrients was launched by the Indian Council of Agricultural Research (ICAR) with headquarters at the Punjab Agricultural University, Ludhiana, which was later shifted to the Indian Institute of Soil Science, Bhopal, Madhya Pradesh. About half of the cultivated soils in the India are de?cient in available zinc. Zinc de?ciency is most in Maharashtra and Karnataka and rice-growing states of north and northeast (Table 12.1). A good response of cereals to Zn fertilization has been reported (Rattan et al., 2008).

There are nine essential micronutrients, namely, iron (Fe), manganese (Mn), zinc (Zn), copper (Cu), boron (B), molybdenum (Mo), chlorine (Cl), vanadium (V) and nickel (Ni). Out of these, Fe, Mn, Zn, Cu and Ni (Dominic et al., 1978) are taken up by plants in their ionic form X²⁺, Mo as MoO^2-, chlorine as Cl^– form and boron as boric acid (H₃BO₃) or as its ionic form B (OH)₄ ^–. The absorption of vanadium by plants has received only limited attention (Lepp et al., 1983); it may be absorbed as cation V or vanadyl (VO²⁺) and anion vanadate (VO₄ ^3–). Bulk of the information is available on Fe, Mn, Zn and Cu, because of the acceptability of the single DTPA (Diethylene triamine pentaacetic acid)–CaCl₂ extraction test of Lindsay and Norvell (1978) for availability of all these four nutrients followed by their estimation on Atomic Absorption Spectrometer (AAS). Available B is estimated by hot 0.15 M CaCl₂ extraction method (Saha and Singh, 1997). Molybdenum data base is poor, because its determination is dif?cult and Mo de?ciency in crops has not been much noticed in India. Chloride is available in plenty in irrigation water and soil in India and has not received much attention in India.

Secondary nutrients is a triad of three essential plant nutrients namely, Calcium (Ca), Magnesium (Mg) and Sulphur (S). Of these three, Ca and Mg are taken up by plants as cations, while S is taken up as anion SO4 2–. Among the secondary plant nutrients, sulphur is emerging as a major plant-nutrient de?ciency in crops grown in the Indo-Gangetic Plains, spread over 13 million ha in Pakistan, India, Nepal, and Bangladesh (Khurana et al., 2008). From the Indian point of view, S de?ciency has emerged in a large part of the country and needs to be attended. Uptake of Mg and S by some crops is given in Table 10.1.

Out of the three primary plant nutrients (NPK), potassium (K) is taken up by crop plants in the largest amount, yet as compared to N and P, only a tiny fraction of it goes in the grain or seed. For example, Shivay et al. (2015) reported from a study on rice that about 20 kg N, 6 kg P (13.7 kg P2O5) and 46 kg K (55.2 kg K2O) was taken up by the crop to produce each Mg or tonne (1,000 kg) of rice kernel; however, only 4.3% of K taken up by the crop was found in the rice kernel as compared to 44% of N and 66% of P taken up by the crop. Some information on N, P and K removed per tonne of grain/ seed for some crops is given in Table 9.1.

As compared to total N soil total P is present in much smaller amounts in soil; it is roughly about 10% of total N. Further, while N in soil is in the largest amount in surface 0 to 15 cm layer, soil P may be the same or in larger amounts in the sub-soil. Again, organic P constitutes only a small portion of total P. Thus, the Chemistry of soil P is generally centered on inorganic P. Total and organic P in some Indian soils is given in Table 8.1; hill soils had the highest and the desert soils the least total as well as organic P.

FRACTIONS OF INORGANIC P

All P in soils has originated from apatites [3 (Ca₃ (PO₄)₂.Ca×₂, where ×₂ could be F, Cl, (OH) or CO₃). Large deposits of apatites form phosphate rocks, which are the major materials for making phosphate fertilizers. As the soil weathering proceeds, P in apatite reacts with Al, Fe and other cations and forms compounds of different solubility. A knowledge of these can help in understanding the P-supplying capacity of soils. A number of chemical extractants and resins are used for determining different fractions. In general, 7 fractions of inorganic soil P are mentioned. These are: saloid-P (soluble and loosely bound P extractable by NH₄Cl), labile P (determined by using anion-exchange resins), Al-P (NH₄F extractable), Fe-P (NaOH extractable), Ca-P (H₂SO₄ or HCl extractable), reductant-soluble- P (CDB or sodium citrate-sodium dithionite-sodium bicarbonate extractable) and occluded P (CTB followed by NaOH extractable)

Fertilizer N has been the key input in increasing cereal production throughout the world. In India, it made possible to achieve the yield potential of high-yielding dwarf varieties of wheat that led to ‘Green Revolution in 1968’. Nitrogen fertilization also increases protein content in wheat grains (Rajeswara Rao and Prasad, 1987) and leads to increased production of proteins in cereals (Prasad and Shivay, 2015). Cereal production in India increased from 42.4 million tonnes (Mt) in 1950–51 to 259.6 Mt in 2017–18, while fertilizer N application increased from 0.05 Mt to 16.96 Mt. However, the average agronomic ef?ciency of N (AEn) in India for cereals is only 9.5 kg grain kg–1 N (Sharma and Batra, 2011) as compared to global average of 24 kg grain kg–1 N for maize, 22 kg grain kg–1 N for rice and 18 kg grain kg–1 N for wheat (Ladha et al., 2005). Besides this, more disturbing is the fact that in irrigated areas the AEn declined from 13.4 to 3.4 kg grain kg–1 N during 1970–2005 (Biswas and Sharma, 2006). This is due to poor N management. Bulk of fertilizer N is applied broadcast on surface in most crops.