CN117121816A - Method for identifying resistance of leaf stalk black spot of Chinese cabbage - Google Patents

Method for identifying resistance of leaf stalk black spot of Chinese cabbage Download PDF

Info

Publication number
CN117121816A
CN117121816A CN202311098392.5A CN202311098392A CN117121816A CN 117121816 A CN117121816 A CN 117121816A CN 202311098392 A CN202311098392 A CN 202311098392A CN 117121816 A CN117121816 A CN 117121816A
Authority
CN
China
Prior art keywords
chinese cabbage
black spot
resistance
culture medium
leaf stalk
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202311098392.5A
Other languages
Chinese (zh)
Inventor
雷娟利
赵彦婷
岳智臣
陶鹏
李必元
胡齐赞
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhejiang Academy of Agricultural Sciences
Original Assignee
Zhejiang Academy of Agricultural Sciences
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Zhejiang Academy of Agricultural Sciences filed Critical Zhejiang Academy of Agricultural Sciences
Priority to CN202311098392.5A priority Critical patent/CN117121816A/en
Publication of CN117121816A publication Critical patent/CN117121816A/en
Pending legal-status Critical Current

Links

Classifications

    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention discloses a method for identifying resistance of leaf stalk black spot of Chinese cabbage, which is characterized in that the resistance of leaf stalk black spot of Chinese cabbage is identified by utilizing a tissue culture mode, and compared with field identification and water culture identification, the method is more time-saving and labor-saving, is not limited by environmental factors, reduces the influence of the environmental factors, and obtains relatively more reliable data.

Description

Method for identifying resistance of leaf stalk black spot of Chinese cabbage
Technical Field
The invention relates to a method for identifying resistance of leaf stalk black spot of Chinese cabbage.
Background
The black spot disease of the leaf stalks of the cabbages seriously affects the quality of the cabbages, reduces the commodity value and causes great loss to producers, operators and consumers. The mechanism responsible for the occurrence of black spot disease in the leaf stalks of chinese cabbage is not completely understood. Previous studies indicate that leaf stalk blackspot of Chinese cabbage is a physiological disease and is the result of the combined action of genotype and environmental factors, and excessive application of N fertilizer is an important factor for causing the disease. The nutrition of nitrogen with different forms also has influence on the occurrence of the black spot disease of the leaf stalks of the Chinese cabbage, wherein the effect of ammonium nitrogen on promoting the occurrence of the black spot disease of the leaf stalks of the Chinese cabbage is higher than that of nitrate nitrogen and amide nitrogen. Gao Xiaorong, and the like, have found that the occurrence of petiole black spot is related to the variety, so that we hopefully solve the problem of petiole black spot from the viewpoint of breeding.
The field identification of the leaf stalk black spot disease of the Chinese cabbage is generally carried out in the harvest period, time and labor are wasted, the period is long, and the method cannot be applied to the breeding work of resisting the leaf stalk black spot disease. Therefore, a set of rapid and effective identification method for the black spot disease of the leaf stalks of the cabbages is established, and the method has extremely important effect on the research and breeding of the black spot disease resistance of the leaf stalks of the cabbages.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method for identifying the resistance of the leaf stalk black spot of the Chinese cabbage, which is used for identifying the resistance of the leaf stalk black spot of the Chinese cabbage, has the characteristics of time and labor saving, is not limited by environmental factors, and has accurate and reliable identification result.
The invention is realized by adopting the following technical scheme:
a method for identifying resistance to leaf stalk black spot of chinese cabbage, comprising the steps of:
(1) Sterilizing 1/2MS culture medium at high temperature and cooling to 50-60 ℃; then adding ammonium nitrate solution to make the concentration of ammonium nitrate in the culture medium be 10mmol/L, and cooling and solidifying the culture medium for later use;
(2) Selecting full Chinese cabbage seeds, sterilizing and preserving;
(3) Sowing the disinfected and sterilized celery cabbage seeds into the solidified culture medium in the step (1), and culturing in a tissue culture chamber with the temperature of 25 ℃ and the illumination intensity of 1200 lx;
(4) After 4-6 weeks of culture, counting the number of black spots to judge the resistance of the leaf stalk black spot of the Chinese cabbage variety.
Preferably, the sterilization temperature in the step is 121 ℃ and the sterilization time is 20min.
Preferably, in the step (2), the Chinese cabbage seeds are sterilized by adopting an ethanol solution with the mass concentration of 75% and a sodium hypochlorite solution with the mass concentration of 10%.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a method for identifying resistance of leaf stalk black spot of Chinese cabbage, which utilizes a tissue culture mode to identify the resistance of leaf stalk black spot of Chinese cabbage, and compared with field identification and water culture identification, the method is more time-saving and labor-saving, is not limited by environmental factors, reduces the influence of the environmental factors, and obtains relatively more reliable data.
Drawings
FIG. 1 is a symptom of black spot disease of petiole of Chinese cabbage in the experimental group and the control group in example 1;
FIG. 2 shows symptoms of leaf stalk black spot of different cabbage varieties in example 3.
Detailed Description
The invention will be further illustrated with reference to specific examples.
Example 1
A method for identifying resistance of leaf stalk black spot of Chinese cabbage comprises the following specific steps:
(1) Sterilizing 1/2MS culture medium at high temperature (121deg.C for 20 min), and cooling to 50deg.C; then adding ammonium nitrate solution to make the concentration of ammonium nitrate in the culture medium be 10mmol/L, pouring the culture medium into a 100mL culture bottle, and cooling and solidifying for later use; taking a culture medium without ammonium nitrate as a control;
(2) Selecting full improved green Chinese cabbage seed No. 3, sterilizing (75 wt% ethanol solution and 10wt% sodium hypochlorite solution) for use;
(3) Sowing the disinfected and sterilized Chinese cabbage seeds in the culture medium (5 seeds are sown in each bottle, 3 plants are reserved in each bottle after the subsequent thinning), and then culturing the culture bottle in a tissue culture chamber with the temperature of 25 ℃ and the illumination intensity of 1200 lx;
(4) After 4 weeks of culture, observing the occurrence of black spots to judge the feasibility of the method;
the results are shown in FIG. 1, wherein FIG. 1 (a) shows an experimental group in which ammonium nitrate was added to the medium, and FIG. 1 (b) shows a control group in which ammonium nitrate was not added to the medium. It can be seen from FIG. 1 that the addition of ammonium nitrate induced petiole black spot in the case of tissue culture.
Example 2
A method for identifying resistance of leaf stalk black spot of Chinese cabbage comprises the following specific steps:
(1) Sterilizing 1/2MS culture medium at 121deg.C for 20min, cooling to 50-60deg.C, respectively adding ammonium nitrate solution and NaCl solution to treat the culture medium, controlling final concentration of both treatments to 10mmol/L, mixing the culture medium, and pouring into 100ml large-sized culture bottle. The culture medium treated by the ammonium nitrate solution is a high-nitrogen group, the culture medium treated by NaCl is a high-salt group, a group of blank control is reserved, and 10 bottles of the culture medium treated by the ammonium nitrate solution and the control group are repeatedly arranged.
(2) Selecting full improved green Chinese cabbage seed No. 3, sterilizing (75 wt% ethanol solution and 10wt% sodium hypochlorite solution) for use;
(3) Sowing the disinfected and sterilized Chinese cabbage seeds in the culture medium (5 seeds are sown in each bottle, 3 plants are reserved in each bottle after the subsequent thinning), and then culturing the culture bottle in a tissue culture chamber with the temperature of 25 ℃ and the illumination intensity of 1200 lx;
(4) After 5 weeks of culture, counting the number of black spots to judge the influence of high N and high salt on the formation of leaf stalk black spots of the Chinese cabbage variety;
the results are shown in Table 1, and indicate that high N is a major factor in the induction of petiole blackspot.
TABLE 1 statistics of small black spots of tissue culture seedlings in different culture media
Example 3
A method for identifying resistance of leaf stalk black spot of Chinese cabbage comprises the following specific steps:
(1) Sterilizing 1/2MS culture medium at high temperature (121deg.C for 20 min), and cooling to 55deg.C; then adding ammonium nitrate solution to make the concentration of ammonium nitrate in the culture medium be 10mmol/L, pouring the culture medium into a 100mL culture bottle, and cooling and solidifying for later use;
(2) Selecting full Chinese cabbage seeds of different varieties, sterilizing (75 wt% ethanol solution and 10wt% sodium hypochlorite solution) for later use;
(3) Sowing the disinfected and sterilized Chinese cabbage seeds in the culture medium (6 seeds are sown in each bottle, 3 plants are reserved in each bottle after the subsequent thinning), and then culturing the culture bottle in a tissue culture chamber with the temperature of 25 ℃ and the illumination intensity of 1200 lx;
(4) After 6 weeks of culture, the number of black spots is counted to judge the resistance of the leaf stalk black spots of different Chinese cabbage varieties.
The results are shown in FIG. 2 and Table 2, which demonstrate that the resistance to leaf stalk black spot is different for different Chinese cabbage varieties.
TABLE 2 statistics of small black spots of tissue culture seedlings of different varieties
It should be noted that the above-mentioned embodiments are only a few specific embodiments of the present invention, and it is obvious that the present invention is not limited to the above embodiments, but other modifications are possible. All modifications directly or indirectly derived from the disclosure of the present invention will be considered to be within the scope of the present invention.

Claims (3)

1. A method for identifying resistance to leaf stalk black spot of chinese cabbage, comprising the steps of:
(1) Sterilizing 1/2MS culture medium at high temperature and cooling to 50-60 ℃; then adding ammonium nitrate solution to make the concentration of ammonium nitrate in the culture medium be 10mmol/L, and cooling and solidifying the culture medium for later use;
(2) Selecting full Chinese cabbage seeds, sterilizing and preserving;
(3) Sowing the disinfected and sterilized celery cabbage seeds into the solidified culture medium in the step (1), and culturing in a tissue culture chamber with the temperature of 25 ℃ and the illumination intensity of 1200 lx;
(4) After 4-6 weeks of culture, counting the number of black spots to judge the resistance of the leaf stalk black spot of the Chinese cabbage variety.
2. The method for identifying resistance to black spot disease of chinese cabbage leafstalk according to claim 1, wherein said sterilizing temperature is 121 ℃ and sterilizing time is 20min.
3. The method for identifying black spot disease resistance of celery cabbage leafstalk according to claim 1, wherein step (2) adopts 75% ethanol solution and 10% sodium hypochlorite solution to disinfect celery cabbage seeds.
CN202311098392.5A 2023-08-29 2023-08-29 Method for identifying resistance of leaf stalk black spot of Chinese cabbage Pending CN117121816A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202311098392.5A CN117121816A (en) 2023-08-29 2023-08-29 Method for identifying resistance of leaf stalk black spot of Chinese cabbage

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202311098392.5A CN117121816A (en) 2023-08-29 2023-08-29 Method for identifying resistance of leaf stalk black spot of Chinese cabbage

Publications (1)

Publication Number Publication Date
CN117121816A true CN117121816A (en) 2023-11-28

Family

ID=88859503

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202311098392.5A Pending CN117121816A (en) 2023-08-29 2023-08-29 Method for identifying resistance of leaf stalk black spot of Chinese cabbage

Country Status (1)

Country Link
CN (1) CN117121816A (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20070002581A (en) * 2005-06-30 2007-01-05 충남대학교산학협력단 Method for regenerating a plant using inbred of chinese cabbage
CN102144561A (en) * 2011-02-25 2011-08-10 西北农林科技大学 Method for in vitro regeneration culture of true leaves of Chinese cabbage
CN102239779A (en) * 2011-03-18 2011-11-16 湖北省农业科学院经济作物研究所 Method for quickly identifying resistance of Chinese cabbage germplasm to black-spot disease
CN105638417A (en) * 2016-02-07 2016-06-08 浙江省农业科学院 Seedling stage screening method for low-calcium-resistant Chinese cabbage strain

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20070002581A (en) * 2005-06-30 2007-01-05 충남대학교산학협력단 Method for regenerating a plant using inbred of chinese cabbage
CN102144561A (en) * 2011-02-25 2011-08-10 西北农林科技大学 Method for in vitro regeneration culture of true leaves of Chinese cabbage
CN102239779A (en) * 2011-03-18 2011-11-16 湖北省农业科学院经济作物研究所 Method for quickly identifying resistance of Chinese cabbage germplasm to black-spot disease
CN105638417A (en) * 2016-02-07 2016-06-08 浙江省农业科学院 Seedling stage screening method for low-calcium-resistant Chinese cabbage strain

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
雷娟利;李必元;岳智臣;陶鹏;赵彦婷;钟新民;: "氮素水平对大白菜叶柄黑点症发生的影响", 浙江农业科学, no. 11, 11 November 2017 (2017-11-11) *
雷娟利等: ""大白菜叶柄黑点症抗性苗期水培鉴定方法"", 《浙江农业科学》, vol. 60, no. 3, 11 March 2019 (2019-03-11), pages 430 - 431 *

Similar Documents

Publication Publication Date Title
Gonbad et al. Influence of cytokinins in combination with GA3 on shoot multiplication and elongation of tea clone Iran 100 (Camellia sinensis (L.) O. Kuntze)
GARDNER BACTERIAL SPOT OF TOMATO¹
CN106106175A (en) Passifolra edulis tissue culture and rapid propagation method
Baum et al. Morphogenetic effect of the herbicide Cinch on Arabidopsis thaliana root development
CN117121816A (en) Method for identifying resistance of leaf stalk black spot of Chinese cabbage
Tiwari Diagnosing potassium deficiency and maximizing fruit crop productivity
CN106718875B (en) Method for growing dendrobium seedlings
Leupin et al. Compact callus induction and plant regeneration of a non-flowering vetiver from Java
CN105815217A (en) Treatment method for bacterial contamination in tissue-culture industrialized seedling culture of tropical woody plants
Jadhav et al. Performance of black gram [Vigna mungo (L.) Hepper] varieties to different sowing dates.
CN107182738A (en) Momordica grosvenori ciltivating process
CN113632675A (en) Cultivation method for improving emergence rate of hybrid progeny of extremely early-maturing pears, pearls and pears
CN106665347A (en) Detoxification and rapid propagation method of sweet potatoes of Yizi 138
CN106359050A (en) Seedling growing method for preventing root-knot nematode disease of fruit cucumbers
Gimenes et al. Forage mass and agronomic traits of Elephant grass genotypes under free-growth and conventional cutting systems.
Wang et al. Construction of tissue culture system of Onobrychis viciaefolia scop ‘Mengnong’anthers
CN1207959C (en) Method of preventing browning in plant tissue culture
CN110393154B (en) Culture method of virus-free tissue culture seedlings of fresh-eating sweet potatoes
CN108521929A (en) Application of the ascorbic acid as termination of diapause accelerating agent in breaking seed dormancy
Pandey et al. Knowledge and adoption of gram production technology
Gupta et al. Breaking seed dormancy in Cleome viscosa L. for improving seed germination
CN107372069A (en) A kind of method for culturing seedlings of Lithops pseudotruncatella
Hoppe et al. The phenological development of three ryegrass clones and the influence of initial silage cut on regrowth under silage management
Rani Factors Responsible for the Yield Gap in Vegetable Cultivation and Measures to Increase the Productivity in Tamil Nadu, India
Chaudhary et al. Effect of growth enhancers as pre harvest spray on flower quality and yield of gladiolus (Gladiolus grandiflorus L.) var. American Beauty

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination