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Method for obtaining crowtoe regeneration plantlet by anther culture

A technology for regenerating plants and L. japonicus, applied in the fields of botanical equipment and methods, plant regeneration, horticultural methods, etc., can solve the problems of self-incompatibility, slow breeding process, poor efficiency, etc. Effects of gene function

Inactive Publication Date: 2013-04-03
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Cultivated japonicus japonicus is autotetraploid, self-incompatible, and has a high degree of heterozygosity. The process of conventional breeding is slow and the efficiency is poor.

Method used

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  • Method for obtaining crowtoe regeneration plantlet by anther culture
  • Method for obtaining crowtoe regeneration plantlet by anther culture
  • Method for obtaining crowtoe regeneration plantlet by anther culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1. The anthers of Lotus corniculatus Linn (Lotus corniculatus Linn) in the uninucleated marginal stage were used as explants, and the regenerated plants of Lotus corniculatus Linn were obtained by in vitro culture.

[0015] The operation process is as figure 1 , the specific process is as follows:

[0016] a. Selection of anthers in the single-nucleus stage of japonicus japonicus japonicus

[0017] 1) The commercially available variety "Leo" of japonicus japonicus was selected as the material. It was sown in early October and started in late April of the following year. Collect about 2mm, green, bracts from 9:00 am to 11:00 am on sunny days. The flower buds are as long as the flower buds and are in the shape of a knife point (use magenta acetate dyeing and pressing method to observe and determine the development stage). Most of the flower buds at this time are in the single-core stage.

[0018] 2) Put the above flower buds into a small kraft paper bag, tak...

Embodiment 2

[0055] The difference between this embodiment and embodiment 1 is:

[0056] ①The formula of callus induction medium was NB+2,4-D1.0mg / L+NAA0.6mg / L+KT2.0mg / L, sucrose 90g / L, agar 6g / L, pH5.8.

[0057] ②The formula of differentiation medium is MS+KT1.0mg / L+NAA0.5mg / L, sucrose 30g / L, agar 6g / L, pH58.

[0058] ③The rooting medium is 1 / 2MS+NAA0.3mg / L, sucrose 10g / L, agar 6g / L, pH5.8.

[0059] ④ Replace the transplanting sterilization matrix with field soil.

[0060] Under the hormone level, the callus induction rate was 29.17%, the differentiation rate was 13.21%, and the rooting rate was 68.89%. The survival rate of transplanting by the method of ④ was 34.21%.

Embodiment 3

[0062] The difference between this embodiment and embodiment 1 is:

[0063] ①The formula of callus induction medium was NB+2,4-D1.5mg / L+NAA0.6mg / L+KT2.0mg / L, sucrose 90g / L, agar 6g / L, pH5.8.

[0064] ②The formula of differentiation medium is MS+KT1.5mg / L+NAA1mg / L, sucrose 30g / L, agar 6g / L, pH5.8.

[0065] ③The rooting medium is 1 / 2MS+NAA0.7mg / L, sucrose 10g / L, agar 6g / L, pH5.8.

[0066] ④ Replace the transplanting sterilized matrix formula transplanting matrix with full nutrient soil.

[0067] Under this hormone level, the callus induction rate was 51.2%, the differentiation rate was 15.68%, and the rooting rate was 55.81%. The survival rate of transplanting by the method of ④ was 60.97%.

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Abstract

The invention relates to a method for obtaining crowtoe regeneration plantlet by anther culture and belongs to the technical field of forage breeding. The method comprises the following steps: (A) obtaining flower buds at the late uninucleate stage, taking the flower buds by an ice box to a lab and refrigerating the flower buds at 4 DEG C for 2-3 days; (B) sterilizing the flower buds by sodium hypochlorite, using an integrated type microscope to isolate the flower buds and placing the isolated substances on a callus induction culture medium for culturing; (C) performing dark culture for 20 days, subculturing for 3-4 times every 15 days, placing the subcultured callus on an adventitious bud culture medium to culture, and inducing the callus to generate a bud; (D) performing rejuvenation culture to the bud, placing the bud on a root medium to culture for 15-20 days, and inducing roots; and (E) hardening the seedling, and transplanting the hardened seedling to obtain the regeneration plantlet. In the operations, steps (B) to (D) are performed under aseptic conditions; the dark culture in the step (C) is performed in a biochemical incubator; and the generation of the adventitious bud in the step (C) and the roots in the step (D) is performed in a manual climatic box.

Description

technical field [0001] The invention relates to a method for obtaining regenerated plants of Lotus corniculatus Linn through anther culture, and belongs to the technical field of forage breeding. Background technique [0002] Lotus corniculatus Linn (Lotus corniculatus Linn) is native to Europe and sub-warm regions, and is a perennial herb of the genus Lotus corniculatus of the family Fabaceae. It has the advantages of high grass yield, long green period, high protein content (crude white matter content in the bud stage is 26.15%), rich nutrition, good palatability and no bloating after livestock feed. Lotus japonicus has the characteristics of heat resistance, flood resistance, barren resistance and acid resistance, and is a promising fine forage grass in warm and humid areas in my country. Therefore, it is of great significance to carry out the breeding research of japonicus japonicus for the development of grassland animal husbandry in the south. [0003] The cultivated...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 魏臻武武自念郑曦陈斐朱平华
Owner YANGZHOU UNIV
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