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Efficient leguminous plant somatic embryogenesis and plant building method

A plant and plant regeneration technology, applied in the fields of plant biology and plant tissue engineering, can solve the problems of low frequency of genetic transformation, escape of foreign genes, high frequency of chimera, etc.

Inactive Publication Date: 2013-08-21
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still many problems, such as the low frequency of genetic transformation (average transformation efficiency is much lower than 2%), high frequency of chimera, and escape of exogenous genes in the widely used soybean genetic transformation system. These problems are seriously delayed. Slow down the genetic breeding and variety improvement process of soybean
[0005] At present, there is no large-scale and long-span molecular breeding method in this field. Therefore, there is a great need in this field to develop new soybean regeneration systems and methods, especially methods for efficient regeneration of soybean plants, so as to overcome the bottleneck problem existing in the prior art

Method used

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  • Efficient leguminous plant somatic embryogenesis and plant building method

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preparation example Construction

[0090] The present invention provides a kind of preparation method of bean somatic embryo, in a preferred example, described method comprises the steps:

[0091] (1) providing cotyledon cut pieces of bean plants, and inducing the cotyledon cut pieces to obtain embryogenic callus; and (2) inducing the obtained embryogenic callus to obtain somatic embryos.

[0092] In another optional example, the cut circumference of the cotyledons accounts for at least 50%, preferably 60%, and more preferably 100% of the total circumference; The cotyledon cutting of 5mm, preferably the cotyledon cutting that the upper surface area is 2mm * 4mm; Preferably 3-5 weeks) when picking.

[0093] In another preference, the induction described in step (1) is to inoculate cotyledon cuttings into MSB culture medium containing 2,4-D and sucrose; the concentration of 2,4-D is 10-50mg / L , preferably 30-40mg / L, most preferably 40mg / L; the concentration of sucrose is 1-10wt%, preferably 5wt%; the light inte...

Embodiment 1

[0122] The preparation of embodiment 1 culture medium

[0123] MSB basal medium is a mixture of salts of MS medium and organic components of B5 medium. MSB mother liquor preparation (1L):

[0124] 700ml of distilled water and then add the following ingredients:

[0125] Macronutrient 10 times 100ml; Calcium chloride 20 times 50ml; Trace element 100 times 10ml; Iron salt 100 times 10ml; B5 organic 100 times 10ml; add 30g sucrose to constant volume;

[0126] Adjust the pH to 5.8; add 8g of agar, boil twice; seal after aliquoting; sterilize at 121°C for 20 minutes.

[0127] The whole process of somatic embryogenesis uses this medium as the basic medium.

Embodiment 2

[0128] Example 2 Get pods

[0129] Select plump and robust soybean seeds and sow them in the field or in the greenhouse in good time. Pick the young pods when the plants bloom for 3-5 weeks, wash them with running tap water for 60 minutes, and absorb the moisture on the surface of the pods. The obtained pods can be stored in a refrigerator at 4°C Backup or direct access to somatic embryo induction.

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Abstract

The invention relates to an efficient leguminous plant somatic embryogenesis and plant building method, and concretely relates to a method for successfully building a large amount of regenerated plants through a somatic cell embryogenesis way of an initial explant which is leguminous tender cotyledon blocks. The method comprises the following steps: peeling and cutting leguminous tender cotyledons to obtain cotyledon blocks; inducing the cotyledon blocks to obtain embryonic calluses; further inducing the embryonic calluses to obtain somatic cell embryos; and regenerating the somatic cell embryos to obtain plants. The leguminous plant somatic embryogenesis method is rapid and efficient, and the genetic stability of the obtained plants is good.

Description

technical field [0001] The invention relates to the fields of plant biology and plant tissue engineering, in particular, the invention relates to high-efficiency somatic embryogenesis and plant establishment of bean plants. Background technique [0002] At present, the global population is expanding, and the earth's resources are increasingly threatened. Food production and food security have become important issues in the 21st century. In my country, the five major crops including rice, wheat, corn, cotton and soybeans are important strategic materials to ensure national economic development and agricultural security. [0003] Beans are the most common economic crops in my country, including soybeans, peas, broad beans, mung beans, red beans, kidney beans, kidney beans, etc. Soybean (Glycine max L.) is a diploid (2n=40) plant belonging to the family Fabaceae, Papilionae, and the genus Soybean. It is not only an important oil crop and food crop, but also the main source of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 朱木兰卫志明
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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