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Rejuvenation and rooting culturing method for genetic transformation seedlings of Jatropha curcas

A technology of genetic transformation and cultivation method, which is applied in the field of rejuvenation and rooting cultivation, can solve the problems of late Jatropha curcas and low rooting rate of transformed seedlings, achieve the effect of thick stems, reduce identification work, and avoid escape

Inactive Publication Date: 2014-04-16
SINO FOREST CHINA INVESTMENTS LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The research work of Jatropha curcas started relatively late, and there are few reports about Jatropha curcas tissue culture and genetic transformation technology, and the rooting rate of transformed seedlings is generally low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0035] In this example, Agrobacterium tumefaciens GV3101 mediated GUS gene transfer into Yunnan Yuanmou wild Jatropha for cultivation. The specific process steps are as follows:

[0036] The preparation step is to infect the explants with Agrobacterium tumefaciens GV3101 containing the target gene vector pCAMBIA1301, and then culture them on a resistant callus induction and differentiation medium containing 2 mg / L glufosinate to obtain regenerated shoots. Sex bud spare;

[0037] In the step of cultivating strong seedlings, the resistant buds are carefully cut from the resistant callus, the browned parts are cut off, and transferred to the strong seedling medium P for strong seedling cultivation. The formula of the strong seedling medium P is: MS basic medium + 30g / L sucrose + 0.2 mg / L 6-benzylaminopurine + 0.005 mg / L indole butyric acid + 1 mg / L silver nitrate + 200 mL / L coconut juice + 7 g / L agar + 2 mg / L glufosinate + 50 mg / L timentin, pH=5.6-6.0, culture conditions are: tempera...

example 2

[0041] In this example, Agrobacterium tumefaciens EHA105 is mediated to transfer GUS gene into wild Jatropha curcas in Yuanmou, Yunnan for cultivation. The specific process steps are as follows:

[0042] The preparation step is to select suitable explants and infect them with Agrobacterium tumefaciens EHA105 containing the target gene vector pBI121-Hyg, and then culture them on a resistant callus induction and differentiation medium containing 5 mg / L hygromycin to obtain regeneration Buds serve as resistant buds for backup;

[0043] In the step of cultivating strong seedlings, the resistant buds are carefully cut from the resistant callus, the browned parts are cut off, and transferred to strong seedling medium P for strong seedling cultivation. The formula of strong seedling medium P is: MS Basic Medium + 30g / L sucrose + 1.5 mg / L 6-benzylaminopurine + 0.1 mg / L indole butyric acid + 5 mg / L silver nitrate + 200 mL / L coconut juice + 7 g / L agar + 5 mg / L hygromycin + 150 mg / L timenti...

example 3

[0047] In this example, Agrobacterium tumefaciens MP90 is used to transfer the FT gene into wild Jatropha spp. in Yuanmou, Yunnan for cultivation. The specific process steps are as follows:

[0048] The preparation step is to select suitable explants and infect them with Agrobacterium tumefaciens MP90 containing the target gene vector pBI121-FT, and then regenerate them on a resistant callus induction and differentiation medium containing 20 mg / L kanamycin Buds serve as resistant buds for backup;

[0049] In the step of cultivating strong seedlings, the resistant buds are carefully cut from the resistant callus, the browned parts are cut off, and transferred to strong seedling medium P for strong seedling cultivation. The formula of strong seedling medium P is: MS Basic Medium + 30g / L sucrose + 1.0 mg / L 6-benzylaminopurine + 0.05 mg / L indole butyric acid + 3 mg / L silver nitrate + 200 mL / L coconut juice + 7 g / L agar + 20 mg / L kanamycin + 100 mg / L timentin, pH=5.6-6.0, culture cond...

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Abstract

The invention relates to a rejuvenation and rooting culturing method for genetic transformation seedlings of Jatropha curcas, which comprises the following steps of: a preparation step of carrying out impregnation treatment on an explant by Agrobacterium tumefaciens, then carrying out kanamycin-resistant callus induction and culturing by a differential medium to obtain regeneration buds serving as resistant buds for later use; a good seedling culturing step of cutting down the resistant buds from kanamycin-resistant callus tissues, cutting off browning positions and transferring the obtained product into a good seedling culture medium P to carry out good seedling cultivation; a rooting culturing step of selecting the resistant buds which is 2 to 3cm higher after being subjected to good seedling cultivation and respectively have 4 to 6 leaves, cutting off the callus tissues at the roots of the resistant buds, soaking the resistant buds in rooting powder solution with the concentration of 10 to 20mg / L for 8 to 15 minutes, and transferring the resistant buds into a rooting culture medium R to carry out rooting cultivation until rooted seedlings reach the technical requirements for taking out of a tank and exercising; and a seedling exercising step of opening a cover to exercise the rooted seedlings which reach the technical requirements for taking out of the tank and exercising and transplanting the rooted seedlings into a greenhouse. According to the method disclosed by the invention, the delicate growth state of the genetic transformation seedlings of Jatropha curcas can be obviously improved, the rooting rate is improved and a good foundation is laid for molecular breeding of Jatropha curcas.

Description

Technical field [0001] The invention relates to the technical field of Jatropha curcas plant genetic transformation, in particular to a method for rejuvenation and rooting of Jatropha curcas genetically transformed seedlings. Background technique [0002] Jatropha curcas, a perennial woody oil plant, belongs to the genus Euphorbiaceae Jatropha, native to America and widely distributed in tropical and subtropical regions. Jatropha curcas is drought-tolerant and barren, and its seed oil content is as high as 60%. It is currently one of the important bioenergy plants. [0003] However, currently Jatropha has the problems of narrow planting area and low yield. However, the traditional breeding cycle is long, and it takes 8-10 years to obtain a variety with excellent traits and stable inheritance. In recent years, the rapid development of plant genetic engineering technology has played an important role in the germplasm improvement of many food and cash crops. Therefore, it has become...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 许文钊孙怀娟王梅珍潘文欢李艳梅陈小莲李耿光
Owner SINO FOREST CHINA INVESTMENTS LTD
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