A method for inducing mature embryo callus of Drunken Horseweed and establishing a regeneration system
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A technology of callus induction and establishment method, which is applied in the directions of plant regeneration, horticultural methods, botanical equipment and methods, etc., and achieves the effect of simple operation and low cost
Active Publication Date: 2018-04-24
LANZHOU UNIVERSITY
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At present, there are few reports on tissue culture using mature embryos of Drunken Horse Grass as explants, so it is necessary to establish a tissue culture regeneration system using mature Drunken Horse Grass embryos as explants. Foundation
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Embodiment 1
[0057] Method concrete steps of the present invention are as follows:
[0058] (1) Selection and treatment of plant materials: Select mature seeds of Drunken Horseweed with uniform color and size, remove the lemma and rinse under running water for 30 minutes, then treat with 45% alcohol for 5 minutes, rinse with sterile water 2-3 times, and use Disinfect with 10% sodium hypochlorite for 10 minutes, rinse with sterile water for 3-5 times, place the seeds on sterile filter paper until the seeds are dry and ready for use; the seeds of Drunken Horse Grass refer to: wild Drunken Horse collected in Xiahe grass seeds.
[0059] (2) The formulations of induction medium, differentiation proliferation medium and rooting medium are as follows:
[0060] The basal medium was MS medium plus 30 g sucrose L -1 and agar 10 g·L -1 ;
[0061] Induction medium and differentiation and proliferation medium were supplemented with different treatment hormones as basal medium.
[0062] The callus ...
Embodiment 2
[0069] The difference between this embodiment and embodiment 1 is:
[0070] The selected Drunken Horse Grass seeds were collected from Tianzhu.
[0071] The basal medium was MS medium plus 30 g sucrose L -1 and agar 10 g·L -1 ;
[0072] The callus induction medium is: basal medium + 0.5 mg·L -1 6-BA+3.0 mg·L -1 2,4-D, the induction rate is 96%;
[0073] The proliferation medium is: basal medium+3.0mg / L6-BA+0.1mg / L NAA, the proliferation rate is 83%;
[0074] The rooting medium is: 1 / 2 basal medium+0.2 mg / L NAA, and the rooting rate is 100%.
Embodiment 3
[0076] The difference between this embodiment and embodiment 1 is:
[0077] The selected seeds of Drunken Horse Grass are the seeds of Drunken Horse Grass collected in Xinjiang.
[0078] The basal medium was MS medium plus 30 g sucrose L -1 and agar 10 g·L -1 ;
[0079] The callus induction medium is: basal medium + 0.5 mg·L -1 6-BA+1.0mg·L -1 2,4-D, the induction rate is 95%;
[0080] The proliferation medium is: basal medium+1.5mg / L6-BA+0.2mg / L NAA, the proliferation rate is 88%;
[0081] The rooting medium is: 1 / 2 basal medium + 0.4 mg / L NAA, and the rooting rate is 100%.
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Abstract
The invention provides an establishment method of an achnatherum inebrians mature embryo callus induction and regeneration system. The establishment method comprises the following steps: (1) selecting achnatherum inebrians seeds and disinfecting; (2) screening an induction culture medium, a differentiation and proliferation culture medium and a rooting culture medium; (3) carrying out treatment and induced differentiation on mature embryos: taking the disinfected achnatherum inebrians seeds of the step (1), and culturing the seeds in the induction culture medium, the differentiation and proliferation culture medium and the rooting culture medium in sequence; after roots of differentiated seedlings grow for 4cm-6cm, moving a seedling bottle out and closing the bottle to carry out seedling hardening; injecting tap water after 2 days; opening the bottle to carry out the seedling hardening for 3 days; washing the roots and sterilizing; and culturing by utilizing a matrix to obtain complete regenerated plants. The achnatherum inebrians mature embryos are used as explants and are subjected to tissue regeneration culture, and the establishment method is not influenced by material taking space-time limitation. The method provided by the invention is simple to operate and low in cost; and the established achnatherum inebrians mature embryo callus induction and regeneration system can provide important basic information for genetic transformation and characteristic improvement.
Description
technical field [0001] The invention belongs to the field of plant regeneration through tissue culture technology in agricultural cultivation, and specifically relates to a method for inducing callus of mature embryos of Drunken Horseweed and establishing a regeneration system. Background technique [0002] Drunken horse grass is a perennial plant of Poaceae and Achnatherum splendens. It is one of the main poisonous weeds in the grasslands of northern my country and is widely distributed in Gansu, Xinjiang, Inner Mongolia, Qinghai and other provinces and regions of China (Shi Zhicheng, 1997). Drunken horsegrass has the advantages of high cold resistance and drought tolerance, and more livestock do not eat it, so it has a greater advantage in interspecific competition in degraded grassland communities, and its area continues to expand, especially in arid and degraded grasslands. In some provinces (regions), Drunken Horse Grass has become the dominant population (Li Xuesen, 19...
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Application Information
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