Some standard content:
ICS65.020.01
Agricultural Industry Standard of the People's Republic of China
NY/T 1281--2007
Rules for Detection of Fungal Diseases on Ornamental Plants2007-04-17 Issued
Implemented on 2007-07-01
Issued by the Ministry of Agriculture of the People's Republic of China
Appendix A and Appendix B of this standard are normative appendices. This standard is proposed and managed by the Ministry of Agriculture of the People's Republic of China. NY/T1281-2007
This standard was drafted by the Quality Supervision, Inspection and Testing Center for Flower Products of the Ministry of Agriculture (Shanghai). The Quality Inspection Center for Vegetables and Flowers of the Ministry of Agriculture (Beijing) participated in the drafting.
The main drafters of this standard are: Lin Dawei, Dai Yongmei, Zhai Jianzhong, Qi Longjun, Sun Qiang, Tang Xiangning, Yang Yuhong, and Liu Su. I
1 Scope
Detection Procedure for Fungal Diseases of Flower Plants
This standard specifies the detection methods for common fungal diseases of major flower plants. This standard is applicable to the detection of common fungal diseases of major flower plants. 2 Normative References
NY/T1281—2007
The clauses in the following documents become the clauses of this standard through reference in this standard. For any dated referenced document, all subsequent amendments (excluding errata) or revisions are not applicable to this standard. However, the parties to an agreement based on this standard are encouraged to study whether the latest versions of these documents can be used. For any undated referenced document, the latest version shall apply to this standard. GB/T18247—2000 Product Grades of Major Flower Plants 3 Terms and Definitions
The following terms and definitions apply to this standard. 3.1
Pathogenspathogen
Parasitic fungi that cause plant diseases, referred to as pathogenic fungi in this article. 3.2
Symptom
After the plant is attacked by pathogenic bacteria, its internal physiological activities and external growth and development show some abnormal state. 3.3
Symptoms
Visible reproductive bodies and vegetative bodies produced by pathogenic bacteria in the affected parts of plants. 3.4
Hypha and mycelium The typical vegetative bodies of fungi are usually filamentous. The single roots are called hyphae, and the aggregate of hyphae is called mycelium. 3.5
Fruitbody
The structure of fungi that produces spores, such as ascocarps, basidiocarps, conidiophores, conidiophores, and stroma, are collectively called fruiting bodies. 3.6
Sclerotium and chlamydospore The structures formed by fungi to resist adverse environments. The hard dormant body formed by the aggregation of hyphae and the thickening of the outer cell wall is called sclerotium. The dormant body formed by the swelling of individual cells and thickening of cell walls in hyphae and spores is called thick-walled spore. 3.7
Colony
The group organization of vegetative or reproductive bodies formed by fungi after they reproduce and expand freely on the surface of artificial culture medium. The colonies of different fungi have specific shapes, sizes and colors.
Asexual spore
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The reproductive body formed directly from the vegetative body by fungi without the combination of sex cells or sex organs, mainly including zoospores, sporangiospores and conidia.
Sexual spore
The reproductive body produced by the combination of two matable sex cells or sex organs of fungi, mainly including oospores, zygospores, ascospores and basidiospores.
4 Sampling
4.1 When testing the quality of flower and plant products by grading, sampling for fungal disease testing shall be carried out in accordance with the following provisions. 4.1.1 For fungal disease testing of fresh cut flowers, sampling shall be carried out in accordance with the provisions of GB/T18247.1-2000. 4.1.2 For fungal disease testing of potted flowers, sampling shall be carried out in accordance with the provisions of GB/T18247.2-2000. 4.1.3 For fungal disease testing of potted foliage plants, sampling shall be carried out in accordance with the provisions of GB/T18247.3-2000. 4.2 Sampling of various flower seedlings, flower bulbs and flowers during the field growth period. Random sampling is adopted, and the number of samples required for sampling is shown in Table 1. Table 1 Sampling Standards for Fungal Disease Detection of Flower Seedlings, Bulbs and Field Plants Total Number of Plants/Bulbs
(Plants/Bulbs)
Below 100
101~300
301~500
501~1000
1001-2000
2.001~5.000
5001 and above
Sampling Percentage
The number of samples in each batch shall not be less than 20 plants (bulbs). If the total number is less than 20 plants (bulbs), all of them shall be inspected. 4.2.1 When flower plants have no obvious symptoms of fungal diseases, sampling shall be carried out according to the smallest proportion in the table. 4.2.2 When flower plants (seedlings, bulbs) are found to have suspicious or obvious symptoms of fungal diseases, sampling shall be carried out according to the larger proportion in the table.
4.3 Sample Storage
4.3.1 Samples tested on the same day: Pack the samples in fresh-keeping bags, seal them, and bring them to the laboratory for testing. 4.3.2 Samples that cannot be tested on the same day: The samples should be wrapped in clean absorbent paper, packed in fresh-keeping bags, sealed, and stored at 4℃～5℃. The shelf life is generally 3d~4d.
5 Main detection objects
5.1 Gray mold of flower plants
Botrytis spp.
5.2 Powdery mildew of flower plants
Oidium spp.
5.3 Anthracnose of flower plants
Anthracnose (Colletotrichum spp.)2
5.4 Penicillium disease of flower plants
Penicillium spp.
5.5 Rust disease of flower plants
Puccinia spp.）
Uromyces pp.
Phragmidium spp.
5.6Leaf spot disease of flower plants
Fulvia spp.
Cladosporium spp.Alternaria spp.
Curuularia spp.
Cercospora spp.
Actinonema spp.
Septoriaspp.
Phyllosticta spp.5.7Root rot disease of flower plants
Rhizoctomia spp.）
Sclerotium spp.
5.8 Blight of Flowers and Plants
Phytophthora spp.
5.9 Wilt of Flowers and Plants (Blight, Root Rot) Fusarium spp.
6 Detection Method
According to the symptom observation, follow the steps listed in Appendix A. 6.2
The laboratory test of pathogenic bacteria shall be carried out according to the steps listed in Appendix B. 7 Result Determination
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7.1 If the results obtained from the symptom observation are consistent with the typical symptoms of a disease in Appendix A, it can be determined as that type of disease. 7.2 If the symptoms cannot be determined, or the pathogen needs to be confirmed, it is necessary to test according to the detection method in Appendix B, and compare the pathogen characteristics obtained by microscopic examination with the pathogen characteristics in Appendix B, and finally make a judgment. 3
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A.1 Symptom test method
A.1.1 Visual observation
Appendix A
(Normative Appendix)
Symptom test of fungal diseases of flower plants
Carefully take the diseased organs (leaves, branches, flowers, roots, etc.) with obvious symptoms, and observe the shape, size, color, texture, etc. of the lesions with the naked eye. A.1.2 Magnifying glass inspection
After visual observation, use a handheld magnifying glass to continue to check the symptoms of the diseased plants, check the presence and characteristics of the symptoms, and determine the type and pathogen of the disease.
A.2 Symptoms and characteristics of major fungal diseases of flower plantsA.2.1: Gray mold of flower plants
It harms the leaves, stems, flowers and fruits of flower plants. In the early stage, water-soaked spots appear, and the diseased tissue gradually becomes soft, yellow or brown and rots. In the later stage, a gray velvety mold layer appears on the spots.
A.2.2 Powdery mildew of flower plants
It mainly harms the leaves, petioles, tender shoots and floral organs of flower plants. A white powdery mold layer grows on the affected parts. Young leaves are infected, and the leaves shrink, thicken and curl; leaves are infected during the growth period, and chlorotic yellow spots appear on the leaf surface, which gradually expand until the whole leaf turns yellow and falls off. The petioles and tender shoots are infected, and the diseased parts are slightly swollen and bent downward, and the internodes are shortened. Flowers are infected, and the mild ones are deformed, and the severe ones wither and lose their ornamental value. Small gray-black particles can be seen in the diseased parts in the later stage.
A.2.3 Anthracnose of Flower Plants
It harms all parts of the plant above ground, causing leaf blight, leaf spots, branch blight, flower rot, fruit spots and fruit rot, etc. The lesions on the leaves are irregular or nearly circular, light brown in the center, brown to dark brown on the edges, with concentric lines, and orange-red or black dots under humid conditions. A.2.4 Penicillium of Flower Plants
It mainly harms the corms, bulbs, tubers and fruits of plants, causing these organs to rot, and later a green powdery mold layer appears on the rotten parts.
A.2.5 Rust of Flower Plants
It mainly harms the leaves, petioles, young branches and buds of flower plants, and sometimes also harms flower buds or young branches. Small yellow spots appear on the front of the affected leaves, and small yellow spots on the back at first, and then turn into dark brown blister-like protrusions. After the blisters burst, yellow, orange-red or brown powdery substances are produced, causing the leaves to fall prematurely.
A.2.6 Leaf spot disease of flowers and plants
Table A.1 Symptoms and characteristics of leaf spot disease of flowers and plants and common symptoms of its pathogens
White or
purple-gray, black-brown, velvety,
tobaccoberry-like or felt-like mold layer appears on the diseased part of the leaves.
Other symptom characteristics
In the early stage, yellow chlorotic spots with unclear edges appear on the front of the leaves, and the back is densely covered with gray-white velvety layers, which later turn purple-gray
The affected leaves often present small round, oval or polygonal spots. The diseased parts fade in the early stage and turn yellow to light black in the later stage. The leaves are infected and produce yellow spots. There are grass-green or brown-green halos around the spots. The spots often heal into pieces, which can cause the death of the entire plant. Pathogenic fungi
Fulvia spp.
Cladosporium spp
Cladosporium spp
Currularia spp.
Common symptoms
White or
purple-grey, dark brown, velvety,
smoky or felt-like layers appear on the diseased parts of leaves.
Small red spots appear on the diseased parts of leaves, which are warty protrusions.
A.2.7 Root rot of flowers and plants
Table A.1 (continued)
Other symptoms and characteristics
The lesions on the affected leaves are round, nearly round, oval or irregular, with ring patterns and dark color.
The lesions on the affected leaves are mostly round, gray-brown, with dark brown edges. Small brown spots appear on the front of the infected leaves, gradually expanding into round, nearly or irregular spots, black-purple, with radial edges, causing leaf fall when the disease is severe. In the early stage, the infected leaves have light yellow to purple spots of varying sizes, gradually expanding into round, oval or irregular brown spots. When the leaves are infected, dead spots gradually form. The spots are round or irregular, yellow-brown to dark brown, and later turn gray-brown to gray in the center, with a clear boundary between the diseased and healthy parts. This refers to the symptoms of root cortical rot caused by fungi. NY/T1281-2007
Pathogenic fungi
Alternaria
Alternariaspp
Cerocospora spp.
Actinomnema spp.
Septoriu spp.
Phyllosticta spp.
Rhizoctonia root rot: The base of the stem of flower seedlings is damaged and rots like water, slightly constricted, and the diseased part is often attached with small soil-like brown sclerotia. Then the root rots, causing the seedlings to wither and die. The seedlings are prone to disease about one month after they emerge from the soil. White rot: Mainly damages the roots. In the early stage, a white filamentous layer appears on the taproot from the base of the stem to the surface soil layer. The root is wet and rotten, and it is easy to pull out of the soil. In the later stage, the plant wilts and dies. When the weather is humid, there are often white mycelium and mouse-feces-like sclerotia at the base of the stem of the diseased plant. A.2.8 Flower and Plant Blight
Mainly harms the leaves of flower plants, but can also harm flowers, stems and roots. Small oily spots appear on the leaves, which gradually expand and are gray-green. When it is humid, a cotton-like mold layer appears on the diseased part, and there is white powder on the mold layer. When the disease is severe, it causes soft rot of leaves and flowers, bending and drooping of stems, and root rot. A.2.9 Flower and Plant Wilt
In the early stage of the disease, the plant tip grows slowly and one side wilts significantly. Young plants are often deformed when affected; when the root is affected, the plant shows changes in leaf color, which are light green, withered yellow, and finally withered grass color. When the diseased stem is cut longitudinally, brown stripes can be seen in the vascular bundle, extending from the root to the tip of the stem.
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B.1 Microscope inspection
B.1.1 Instruments and equipment
Appendix B
(Normative appendix)
Laboratory inspection methods for fungal diseases of flowers and plants High-power biological microscope, solid biological microscope. B.1.2 Reagent preparation
B.1.2.1 Glycerol lactic acid
Distilled water
B.1.2.2 Alcohol and glacial acetic acid mixture
Glacial acetic acid
95% alcohol
B.1.2.3 Aniline blue aqueous solution
Aniline blue
Distilled water
B.1.2.4 Chloral hydrate aqueous solution
5g Chloral hydrate
Distilled water
B.1.3 Inspection method
B.1.3.1 Picking of mycelium and fruiting bodies For microscopic examination, if there are a large number of mycelium or fruiting bodies on the specimen, they can be picked directly with a needle, placed on a slide with a drop of floating medium (usually distilled water), covered with a coverslip and observed under a microscope. B.1.3.2 Sticking microscopic examination of leaf surface fungi
Botrytis cinerea, various leaf spot pathogens, penicillium or rust pathogens on the leaf surface can be examined by sticking method. The specific method is to stick a small piece of transparent tape on the leaf surface where the fungi grow, and the fungi will stick to the tape. Remove the tape and place it in a drop of glycerol lactic acid on a slide, add another drop of glycerol lactic acid on it, and observe with a cover glass. B.1.3.3 Whole tissue transparent microscopic examinationbZxz.net
Put a small piece of diseased tissue slice in a mixture of alcohol and glacial acetic acid for 24 hours, then immerse it in a saturated chloral hydrate aqueous solution, take it out and wash it with water after the tissue is transparent, then stain it with aniline blue aqueous solution, then put it in glycerol lactic acid float and observe it under a microscope. B.1.3.4 Freehand section microscopic examination
Freehand sectioning is performed with a razor or a single-sided blade. If the material is relatively hard and coarse, cut it directly between the fingers; small and soft materials can be cut between the pith, fresh carrots or potato pieces. First trim the material to a suitable size (1cm to 2cm long, about 0.5cm in diameter), hold it with the left index finger and thumb, and hold the lower end of the material with the middle finger, so that the material protrudes 1mm to 2mm above the finger. Hold the knife in the right hand and cut it diagonally from left to right. When cutting, make the blade perpendicular to the material. After cutting 4 to 5 thin slices, use a brush dipped in water to gently remove them along the blade and put them into the culture blood with distilled water. When a certain number of slices are cut, select the thin slice material with pathogens in the culture III for microscopic examination. You can also soak the diseased tissue, place it on a small wooden block with a very flat surface, load a glass slide on it and gently press it with your fingers. As the fingers slowly move backward, use force to pull the material into thin slices. B.2 Pathogen culture test
B.2.1 Instruments and equipment
Autoclave, constant temperature incubator, clean bench. B.2.2 Reagent preparation
B.2.2.1 PDA culture medium
Potato
Glucose
17g~20g
1000mL
B.2.2.2 Oatmeal agar culture medium
Oatmeal
17g~20g
1000mL
B.2.2.3 3%~5% sodium hypochlorite aqueous solution
Sodium hypochlorite 3mL~5mL
Distilled water
Add to 100mL
B.2.3 Moisture culture detection method
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Put 2 filter papers in a culture dish, moisten them with sterile water, and put two thin glass rods on them. Rinse the suspicious specimens with sterile water and place them on the glass rods. Cultivate them at 20℃～25℃ for a certain period of time. When the specimens grow mycelium or fruiting bodies, perform microscopic examination. B.2.4. Separation culture detection method
B.2.4.1 Selection of materials
Select fresh plants, organs or tissues with typical symptoms, wash them, dry them, and cut the border between the diseased and healthy parts into 3mm～5mm square pieces for use as separation materials. If the materials have been severely corrupted and cannot be separated and cultured conventionally, the method of inoculation and separation can be used, that is, the diseased tissue is used as the inoculation material, and it is directly (or traumatically) inoculated on healthy plants or in vitro plant materials, and then separated and cultured from the diseased plants or diseased tissues after the disease occurs.
B.2.4.2 Disinfection and sterilization of tools
Open the clean bench for ventilation for more than 20 minutes, wipe hands, countertops and workbench air outlets with 70% alcohol for disinfection, and wipe the containers and tweezers used for separation with 95% alcohol and then sterilize them by flame burning. B.2.4.3 Preparation of flat PDA
Put the PDA culture medium in the triangular bottle to be used in a microwave oven to melt, take out the shaking hook, and pour the culture medium into the sterilized culture III (thickness 2mm3mm) on the clean bench through aseptic operation, shake well, and let it stand on the table to cool. When pouring the culture medium, do not let the mouth of the triangular bottle touch the wall of the culture medium to prevent the culture medium from adhering to the wall of the dish and causing contamination. To prevent bacterial contamination, 3 drops of 25% lactic acid can be added to 10mL of the culture medium, or streptomycin (40μg/mL) can be added to the culture medium.
B.2.4.4 Disinfection of materials
Put the separated materials in a sterilized small container, rinse with 70% alcohol for 2s~3s, quickly pour it out, and then disinfect with 3%~5% sodium hypochlorite aqueous solution for 5min~15min, then rinse with sterile water for 3~4 times, and finally use sterilized filter paper to absorb the water on the materials. B.2.4.5 Transplantation of materials
Use tweezers to transfer the materials to the flat PDA culture medium (oatmeal agar culture medium is used for Phytophthora) under sterile operation. Multiple pieces of separated materials can be placed separately on one culture dish. 7
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B.2.4.6 Fungal culture of materials
Mark the separated materials and date on the culture dish, and culture the culture blood in a constant temperature incubator at about 25℃ in the dark. When obvious fungal mycelium or fruiting fruit grows in the culture dish, it can be identified. B.3 Identification characteristics of flower plant pathogens
B.3.1 Botrytis spp. Conidiophores are clustered, straight or slightly curved, light brown, smooth, with septa, with 1-2 branches at the top, and the ends of the branches are often obviously swollen, forming colorless or light-colored bottle-shaped spore-producing cells, which are mostly bud-producing, with a size of 960μm~1200μm×16μm~22μm. Conidia are oval, unicellular, colorless or gray, with a size of 9um~16μm×6μm~10um, and are gathered in piles and appear gray. The pathogen forms black irregular sclerotia. The sexual state is Sclerotinia or Botryotinia. B.3.2 Powdery mildew fungus (Oidium spp.)
The mycelium is white, superficial, and absorbs nutrients by deep haustoria into the host epidermal cells. A felt-like powdery mildew layer is often produced on the host surface, which is the mycelium, conidiophores and conidia. Conidiophores are erect and unbranched. Conidia are oblong, unicellular, colorless, and grow in strings, maturing in sequence from top to bottom. The sexual state is many genera in the Erysiphaceae family. B.3.3 Colletotrichumspp. Conidia are disc-shaped, growing under the epidermis of the host plant, scattered or fused. Sometimes there are bristles, which are brown, septate, and gradually pointed to the top. Conidiophores are straight, blunt at the top, often colorless, and 9m~24um×3μm~4.5μm in size. Conidia are short cylindrical to crescent-shaped, colorless, unicellular, thin-walled, smooth on the surface, and sometimes contain oil balls. Sclerotia are occasionally produced. The pathogen is differentiated into multiple specialized types based on the differences in pathogenicity to different hosts.
B.3.4 Penicillium spp. Conidiophores are colorless, solitary, with rough or smooth surfaces and septa. A few of them are aggregated into conidiophore bundles, which are upright, with one or more broom-like branches at the top. The branches form a large number of spore-producing cells at the top, on which clusters of conidia are produced, which mature in sequence from top to bottom. Conidia are round or oval, colorless, unicellular, with smooth or warty surfaces, and appear cyan or green when aggregated. B.3.5 Rust fungi
B.3.5.1 Puccinia spp. Winter spores are produced under the epidermis and are mostly exposed. Winter spores are twin cells, sometimes mixed with rare single cells or multiple cells, each cell has a germination hole, the germination hole of the top cell is mostly born at the top, and the germination hole of the next cell is immediately under the septum, the spore wall is colored, smooth or decorated; the stalk is colored or colorless, not gelatinized, permanent or can fall off, germinate to form a septate basidia, and basidiospores are born on the upper epidermis. Summer spores are born under the epidermis and are naked. Summer spores are solitary on the stalk, single-celled, round or oval, with colored spore walls, small thorns, and multiple germination holes. Sexual spores are buried under the upper epidermis and are nearly spherical. Rust spores are born under the lower epidermis, and are cup-shaped or cylindrical when exposed, with a capsule. Rust spores are round or oval, often squeezed together to form polygons, and the spore wall is colorless with small tubercles.
B.3.5.2 The winter spores of the single-celled rust fungus (Uromyces pp.) grow under the epidermis of the host, and are dark brown to black and powdery after rupture. The winter spores of the single-celled rust fungus are stalked, yellow-brown to chestnut, smooth, with a thick top wall and a germination hole at the top. The summer spores grow under the epidermis of the host, and are reddish brown and powdery after exposure. The summer spores are single-celled, nearly round or oval, yellow-brown, with thorns or tubercle-like protrusions on the surface, solitary on the stalk, and the germination hole is obvious. The sexual spores are often buried in the host tissue, with lateral threads protruding from the orifice. The rust spores are cup-shaped or short cylindrical and have a capsule. The rust spores are round to oval, light-colored, smooth or with small tubercles, and grow in strings. B.3.5.3 The winter spores of the multicellular rust fungus (Phragmidiumspp.) grow on the back of the leaves, which are small, round or irregular, black, and often have lateral threads. Winter spores are solitary on the stalk, large and conspicuous, 3 to multiple cells, each cell has 2 to 3 lateral germination holes, thick wall, smooth or with small tubercles; the stalk is long and swollen, often absorbent, the upper part of the stalk is colored, and the lower part is colorless. The summer spores are initially born under the host epidermis, then exposed, orange-yellow, surrounded by lateral threads. Summer spores are round or elliptical, solitary on the stalk, with small spines or tubercles on the wall, and the germination holes are scattered and not very conspicuous. Sexual spores are born under the cuticle or in the epidermis, conical or oblate. Rust spores are initially born under the epidermis, then exposed, round or cylindrical, without a capsule, with lateral threads. Rust spores are round to elliptical, with tubercles, growing in series, and parasitizing on a single host. 8
B.3.6 Leaf spot pathogen
B.3.6.1 Fulvia spp.
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Mycelium is sparse and has stroma. Conidiophores extend from stroma, unbranched or occasionally branched, straight or curved, narrow at the base, gradually widening upwards, with node-like swellings on one side of the stalk, which can grow laterally as short side branches, light brown to dark brown, smooth. Conidia grow in strings, spore chains often branch, spore tops grow laterally, obtuse at both ends, light brown to dark brown, smooth surface, 0 to 3 septa, sometimes with a slightly protruding umbilicus. B.3.6.2 Cladosporium spp. Mycelium is buried or surface, sometimes producing stroma. Conidiophores are dark brown, solitary or clustered, unbranched or only branched in the middle and upper parts. Conidia are solitary or in short chains, dark brown, with great variation in shape and size, oval, cylindrical, lemon-shaped or irregular, with 0 to 3 septa, smooth surface or with small warts, and obvious spore scars and hilums. The sexual state is Mycosphaerella, Venturia or Apiospora.
B.3.6.3 Alternariaspp. Mycelium is completely buried or partially surface. The hyphae are slender, branched, colorless or brown with septa. Rarely form stroma, without bristles. Conidiophores are thicker than hyphae, light brown to brown, single branched or with loose irregular branches, curved or knee-shaped. Conidia are chained or solitary, light brown to dark brown, with different shapes, typically oval or inverted clavate, smooth surface or with slight spines, with longitudinal and transverse septa, and often with beak-shaped cells at the top. Can form long or short, branched or unbranched spore chains. B.3.6.4 Curvularia spp.) The stroma is relatively large, erect, black, cylindrical, and sometimes branched. The conidiophores are brown, solitary or clustered, straight or curved, and often flexed! The surface is smooth and has septa. Conidia are solitary, apical and lateral, unbranched, often curved to one side, clavate, elliptical, broad fusiform, obovate or pear-shaped, with more than 3 septa, light brown to dark brown, usually with lighter cells at both ends, sometimes with dark bands at the septa, smooth surface or with slight thorns; some have prominent umbilicus points, and some form tridentate star-shaped spores. B.3.6.5 Cercospora sp. Mycelium is mostly epiphytic, often with stroma, conidiophores are thick hyphae-shaped, brown, and the top is lighter in color. Unbranched or occasionally branched, straight or curved, sometimes in a knee-like shape, clustered on the stroma tissue. Conidia are solitary, with scars at the base, colorless or light brown, needle-shaped, whip-shaped or oblong, straight or slightly curved, with multiple septa and smooth surface. Most of the sexual forms belong to the genus Mycosphyaerella. B.3.6.6 Actinonemaspp. The conidiophores are surrounded by radial hyphae, which are cushion-like tissues, attached to the epidermis, and break through the epidermis after maturity. The conidiophores are short. Conidia are oblong or gourd-shaped, twin, slightly constricted at the septum, and the upper cells are smaller with beak-like protrusions, which are biased to one side. This genus is similar to Marssonina, which does not form radial hyphae. B.3.6.7 Septoriaspp. The conidiophores are spherical, buried, brown, thin-walled, solitary and rounded, with dark-colored orifices, thickened cell walls, and sometimes papillae. Conidiophores are absent. Conidia are linear, colorless, with multiple septa and smooth surface. Most of the sexual forms found are Myrosphaerlla and Leptosphaeria. B.3.6.8 Phyllosticta spp. Conidia are buried, single-chambered or multi-chambered, scattered or aggregated, dark brown or black brown, with a centrally centered orifice and round. There are two types of conidia: large conidia are nearly oval or elliptical, unicellular, colorless, slightly pointed at the base, with smooth walls, oil balls, terminal G-type appendages, and no branches; small conidia are cylindrical or dumbbell-shaped, unicellular, and colorless. Phyllosticta spp. grows faster on artificial culture media, with more aerial hyphae, sparse formation of conidia, and membranous walls.
B.3.7 Root rot pathogens
B.3.7.1 Rhizoctonia spp. Mycelium is initially colorless, then turns brown, with septa forming near the branches, and is constricted; the mycelium is mostly right-angled and radial. The sclerotia are initially white, then turn brown, or reddish brown, irregular in shape, with a rough surface, growing in the mycelium, and connected to each other by mycelium. B.3.7.2 Sclerotium spp. 9
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Mycelium is white, lush, and expands radially. The mycelium branches are not at right angles and have septa. Sclerotia are spherical or irregular, brown to black, with compact tissues, and a diameter of 0.5mm~3mm
B.3.8 Phytophthora spp. Mycelium is unseptate, well-developed, multi-branched, and 4um~8um in diameter. Some species have locally enlarged hyphae, called mycelial expansion bodies, which are terminal or intercalary, spherical, nearly spherical, elliptical or irregular in shape. Sporangiophores are slightly different from mycelium. Usually, these hyphae are thin, with a diameter of 2μm~4μm. A few species have clearly differentiated sporangiophores, which are unbranched or monoaxially branched, and a few species have complex branches. New sporangia grow from the inner base of old empty sporangia or from the outer side of the base of old sporangia. Multiple sporangia can often be produced on one sporangiophore. Sporangium is terminal, nearly spherical, ovoid, pear-shaped, oblong or irregular, with papillae or no papillae on the top of the neck; papillae-bearing sporangium usually fall off after maturity, and the fallen sporangium has a handle; sporangium without papillae does not fall off. Zoospores are formed in the sporangium and released from the spores at the top during germination; a few sporangium can germinate directly and grow germ tubes. Thick-walled spores are present or absent, spherical, often with thick walls and smooth. The oospore is spherical, with smooth walls, and a few species have bubble-like protrusions, containing one oospore; the anther is lateral or inferior, and the lateral anther is usually attached to one side of the oospore handle, while the inferior anther is wrapped around the oospore handle. Oospores are thick-walled, smooth, and light yellow to yellow-brown. B.3.9 Fusarium spp.
Conidiophores are colorless, with or without septa, and often aggregate into conidiophores at the lower end, sometimes growing directly from hyphae, unbranched or multi-branched, with spore-producing cells at the top. Large conidia are elliptical or sickle-shaped, colorless, multicellular, slightly pointed at both ends, slightly curved, and the base cells often have an obvious protrusion called a foot cell; small conidia are oval to elliptical, colorless, unicellular or bicellular, solitary or in clusters. Some species can form thick-walled spores at the end or middle of the hyphae or on conidia. Thick-walled spores are spherical or nearly spherical, with a smooth surface or serrated protrusions, mostly colorless, a few yellow-brown, solitary, in clusters or aggregated. The sexual state belongs to the genera Gibberella, Nectria, Hypomyces and Calonectia, etc. B.4 Result determination
If the characteristics of the pathogen meet any of the categories in B.3, it can be determined to be that pathogen. 10) The hyphae are unseptate, well-developed, multi-branched, and 4um to 8um in diameter. Some species have locally enlarged hyphae, called hyphal expansion bodies, which are terminal or intercalary, spherical, nearly spherical, elliptical or irregular in shape. The sporangium stalk is slightly different from the hyphae. Usually, this hyphae is thin, with a diameter of 2μm to 4μm. A few species have clearly differentiated sporangium stalks, which are unbranched or monoaxially branched, and a few species have complex branches. New sporangia grow from the inner base of the old empty sporangium or from the outer side of the base of the old sporangium. Multiple sporangia can often be produced on one sporangium stalk. Sporangia are terminal, nearly spherical, ovate, pear-shaped, oblong or irregular in shape, with papillae or no papillae on the top of the neck; sporangia with papillae usually fall off after maturity, and the fallen sporangia have handles; sporangia without papillae do not fall off. Zoospores are formed in the sporangium and released from the spores at the top when germinating; a few sporangia can germinate directly to grow germ tubes. Thick-walled spores are present or absent, spherical, often with thick walls and smooth. The oospore is spherical, with smooth walls mostly, and a few species have bubble-like protrusions, containing one oospore; the anther is lateral or inferior, the lateral anther is usually attached to one side of the oospore handle, and the inferior anther is wrapped around the oospore handle. Oospores are thick-walled, smooth, and light yellow to yellow-brown. B.3.9 Fusarium spp.
Conidiophores are colorless, with or without septa, and the lower end is often aggregated into conidiophores, sometimes directly growing from hyphae, unbranched to multiply branched, with spore-producing cells at the top. Large conidia are elliptical or sickle-shaped, colorless, multicellular, slightly pointed at both ends, slightly curved, and the base cells often have an obvious protrusion called a foot cell; small conidia are oval to elliptical, colorless, unicellular or bicellular, solitary or in clusters. Some species can form thick-walled spores at the end or in the middle of the hyphae or on the conidia. Thick-walled spores are spherical or nearly spherical, with a smooth surface or serrated protrusions, mostly colorless, a few yellow-brown, solitary, in clusters or aggregated. The sexual state belongs to the genera Gibberella, Nectria, Hypomyces and Calonectia, etc. B.4 Result determination
If the characteristics of the pathogen meet one of the categories in B.3, it can be determined as the pathogen. 10) The hyphae are unseptate, well-developed, multi-branched, and 4um to 8um in diameter. Some species have locally enlarged hyphae, called hyphal expansion bodies, which are terminal or intercalary, spherical, nearly spherical, elliptical or irregular in shape. The sporangium stalk is slightly different from the hyphae. Usually, this hyphae is thin, with a diameter of 2μm to 4μm. A few species have clearly differentiated sporangium stalks, which are unbranched or monoaxially branched, and a few species have complex branches. New sporangia grow from the inner base of the old empty sporangium or from the outer side of the base of the old sporangium. Multiple sporangia can often be produced on one sporangium stalk. Sporangia are terminal, nearly spherical, ovate, pear-shaped, oblong or irregular in shape, with papillae or no papillae on the top of the neck; sporangia with papillae usually fall off after maturity, and the fallen sporangia have handles; sporangia without papillae do not fall off. Zoospores are formed in the sporangium and released from the spores at the top when germinating; a few sporangia can germinate directly to grow germ tubes. Thick-walled spores are present or absent, spherical, often with thick walls and smooth. The oospore is spherical, with smooth walls mostly, and a few species have bubble-like protrusions, containing one oospore; the anther is lateral or inferior, the lateral anther is usually attached to one side of the oospore handle, and the inferior anther is wrapped around the oospore handle. Oospores are thick-walled, smooth, and light yellow to yellow-brown. B.3.9 Fusarium spp.
Conidiophores are colorless, with or without septa, and the lower end is often aggregated into conidiophores, sometimes directly growing from hyphae, unbranched to multiply branched, with spore-producing cells at the top. Large conidia are elliptical or sickle-shaped, colorless, multicellular, slightly pointed at both ends, slightly curved, and the base cells often have an obvious protrusion called a foot cell; small conidia are oval to elliptical, colorless, unicellular or bicellular, solitary or in clusters. Some species can form thick-walled spores at the end or in the middle of the hyphae or on the conidia. Thick-walled spores are spherical or nearly spherical, with a smooth surface or serrated protrusions, mostly colorless, a few yellow-brown, solitary, in clusters or aggregated. The sexual state belongs to the genera Gibberella, Nectria, Hypomyces and Calonectia, etc. B.4 Result determination
If the characteristics of the pathogen meet one of the categories in B.3, it can be determined as the pathogen. 10
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