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Method for plant regeneration and somatic embryogenesis of masson pine capable of resisting against pine wilt diseases

A technology for pine wood nematode disease and embryogenesis, which is applied in plant regeneration, botanical equipment and methods, horticultural methods, etc., and can solve the problems of difficulty in obtaining regenerated plants, difficulty in differentiation, and low germination rate.

Pending Publication Date: 2018-01-19
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there have been related studies on somatic embryogenesis of Pinus massoniana, but there are still problems such as low induction rate of embryogenic callus, difficulty in differentiation, low germination rate, and difficulty in obtaining regenerated plants.

Method used

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  • Method for plant regeneration and somatic embryogenesis of masson pine capable of resisting against pine wilt diseases
  • Method for plant regeneration and somatic embryogenesis of masson pine capable of resisting against pine wilt diseases
  • Method for plant regeneration and somatic embryogenesis of masson pine capable of resisting against pine wilt diseases

Examples

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Embodiment 1

[0027] A method for somatic embryogenesis and plant regeneration of Pinus massoniana resistant to pine wood nematode disease, taking the resistant Pinus massoniana family Ma 7-6 as an example, the process is as follows:

[0028] 1) Source and processing of explant material

[0029] From June to August of 2014 to 2015, in Anhui masson pine resistant candidate individual preservation garden, the immature cones of resistant masson pine were collected in batches at fixed points and plants, and 4 excellent trees were determined, and 6 cones were collected from each plant. Collect once a week, and store in a refrigerator at 4°C for 7 days on the day of collection. Referring to the operation method of Li Qingqing et al. [Li Qingqing, Ye Jianren, Zhu Lihua, et al. 2012. Somatic embryogenesis and plant regeneration of immature embryos of black pine. Forestry Science, 48 (12): 39-44], the seeds of Pine massoniana were disinfected, stripped The sterile female gametophytes (ie immature z...

Embodiment 2

[0067] A method for somatic embryogenesis and plant regeneration of Pinus massoniana resistant to pine wood nematode disease, taking the resistant Pinus massoniana family and 15-1 as examples, the process is as follows:

[0068] 1) Treatment of explants and induction of embryogenic callus

[0069]The method is the same as Example 1, and in the optimal medium LP+2mg / L 2,4-D+1mg / L 6-BA, the highest induction rate of embryogenic callus is 13.1%.

[0070] 2) Maintenance and proliferation of embryogenic callus

[0071] The method is the same as Example 1, and the embryogenic callus can proliferate stably. Proliferation culture was carried out in two ways: solid culture and suspension culture.

[0072] 3) Maturation, differentiation and germination of resistant P. massoniana somatic embryos

[0073] The method is the same as Example 1, with LP as the basic medium, adding 2mg / L ABA, 0.5mg / L GA 3 , 130g / LPEG8000, 25g / L maltose, 1g / L activated carbon, 3.5g / L gel maturation medium, ...

Embodiment 3

[0078] A method for somatic embryogenesis and plant regeneration of Pinus massoniana resistant to pine wood nematode disease, taking the resistant Pinus massoniana family Guang 42-1 as an example, the process is as follows:

[0079] 1) Treatment of explants and induction and proliferation of embryogenic callus

[0080] The method is the same as Example 1, and in the optimal medium LP+2mg / L 2,4-D+1mg / L 6-BA, the highest induction rate of embryogenic callus is 6.8%.

[0081] 2) Maintenance and proliferation of embryogenic callus

[0082] The method is the same as Example 1, and the embryogenic callus can proliferate stably. Proliferation culture was carried out in two ways: solid culture and suspension culture.

[0083] 3) Maturation, differentiation and germination of resistant P. massoniana somatic embryos

[0084] The method is the same as in Example 1, taking LP as the basic medium, adding 2mg / L ABA, 0.5mg / L GA 3 , 130g / LPEG8000, 25g / L maltose, 1g / L activated carbon, 3.5...

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Abstract

The invention discloses a method for plant regeneration and somatic embryogenesis of masson pine capable of resisting against pine wilt diseases. The method comprises the following steps: by taking immature zygotic embryo of the masson pine capable of resisting against pine wilt diseases as a material, performing embryonic callus induction, embryonic callus multiplication, maintenance, differentiation, maturation and germination; performing rooting induction on the somatic embryo; performing strong seedling culture and domestication transplanting on the regenerated plant; and lastly, acquiringthe masson pine plant capable of resisting against pine wilt diseases. According to the method for plant regeneration and somatic embryogenesis of masson pine capable of resisting against pine wilt diseases disclosed by the invention, the immature zygotic embryo of the resistant masson pine is taken as the material, a resistant masson pine somatic embryogenesis regeneration system is preliminarily established, the embryonic callus induction for the resistant masson pine is optimized, the important links, such as, somatic embryo multiplication, differentiation, germination and regeneration plant transplanting domestication, are further researched, a small quantity of resistant masson pine suspension culture and somatic embryogenesis regenerating plants are cultivated and an excellent technical basis is established for the industrialized production of the resistant masson pine.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for somatic embryogenesis and plant regeneration of Pinus massoniana resistant to pine wood nematode disease. Background technique [0002] Masson pine is the most widely distributed, widely used, and most utilized native tree species for afforestation in my country among coniferous species (Chen Bihua et al., 2010). Pine wood nematode disease is a devastating pine tree disease caused by the pine wood nematode (Bursaphelenchus xylophilus), which has the characteristics of easy transmission, rapid onset, and difficult control. In recent years, the occurrence of pine wood nematodes in masson pine forests has caused serious damage in some areas. Therefore, breeding excellent pine varieties with resistance to pine wood nematode and improving the quality of pine resistance is one of the effective ways to preserve pine resources and fundamentally resi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 吴小芹陈婷婷叶建仁沈李元
Owner NANJING FORESTRY UNIV
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