Peanut somatic embryo induction and plant regeneration method
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A peanut and plant technology, applied in plant regeneration, botanical equipment and methods, horticultural methods, etc., can solve the problem of low frequency of plant regeneration, and achieve the effect of avoiding chimerism and high regeneration rate.
Inactive Publication Date: 2011-02-09
QINGDAO AGRI UNIV
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[0005] The invention provides a method for induction of somatic embryos of peanuts and regeneration of plants, which can solve the problem of
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Embodiment 1
[0021] A method for induction of peanut somatic embryos and plant regeneration, comprising the steps of:
[0022] 1 Preparation of somatic embryo induction medium:
[0023] Pick MSB 5 (MS inorganic salt+B 5 Organic component) is the basic medium, adding 3% sucrose and 0.8% agar, MSB 5 The pH value of the medium was adjusted to 5.8, and the culture was carried out under the conditions of the temperature of the culture room at 24-26°C, the light intensity of 2500-3000Lx, and the daily light of 13h. In this MSB 5 Add 2,4-D to the medium to form somatic embryo induction medium, and the amount of 2,4-D added is per liter of MSB 5 Add 5 mg to the medium, 2,4-D is a chlorinated phenoxyacetic acid herbicide, which can also be used as a plant growth regulator, and is a common auxin used to induce callus formation.
[0024] 2 Selection of explant material:
[0025] The test materials are Huayu No. 20, Huayu No. 22, Huayu No. 23 and Huayu No. Yu No. 25, and the new peanut variety L...
Embodiment 2
[0036] A method for induction of peanut somatic embryos and plant regeneration, comprising the steps of:
[0037] 1 Preparation of somatic embryo induction medium:
[0038] Pick MSB 5 For basic medium, add 3% sucrose, 0.8% agar, MSB 5 The pH value of the medium was adjusted to 5.8, and the culture was carried out under the conditions of the temperature of the culture room at 24-26°C, the light intensity of 2500-3000Lx, and the daily light of 10-15h. 5 Add 2,4-D to the medium to form somatic embryo induction medium, and the amount of 2,4-D added is per liter of MSB 5 Add 10 mg to the medium.
[0039] 2 Selection of explant material:
[0040]The test materials are Huayu No. 20, Huayu No. 22, Huayu No. 23 and Huayu No. Yu No. 25, and the new peanut variety Luhua No. 11 selected and promoted by Qingdao Agricultural University (formerly Laiyang Agricultural College), and the seed embryos of mature peanut seeds with large and full grains in each variety after removal of cotyled...
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Abstract
The invention provides a peanut somatic embryo induction and plant regeneration method, which can solve the problem of low plant regeneration frequency in the prior art. The method of the invention comprises the following main steps: separating embryonic leaflet from a peanut embryo which is treated by surface sterilization and soaked, and inoculating the embryonic leaflet to an induction culture medium to which 2,4-D (5-10mg/L) is added for culturing a somatic embryo; then transferring the somatic embryo to a somatic embryo germinating culture medium to which BAP (4mg/L) is added, and subculturing twice to obtain regenerated plants; and cutting the somatic embryo at the bottom after the somatic embryo germinates and elongates, and transferring the cut somatic embryo to a 1/2MS inorganic salt and B5 organic rooting culture medium to which NAA (0.2-0.5mg/L) is added for inducing rooting to obtain complete plants. By taking the embryonic leaflet of peanut mature seeds as explant, and using the induction culture medium of 2,4-D (5-10mg/L) and the somatic embryo germinating culture medium of BAP (4mg/L), the somatic embryo induction rate and plant regeneration rate are high, and simultaneously the somatic embryo is derived from one cell, and the chimerism of regenerated plants is avoided.
Description
technical field [0001] The invention relates to a method for cultivating peanuts, in particular to a method for inducing somatic embryos of peanuts and regenerating plants. Background technique [0002] Peanut is an important oil crop and economic crop in my country, and plays an important role in agriculture and even the entire national economy. However, due to the narrow genetic base and poor stress resistance of peanut cultivars, they are especially susceptible to various diseases and insect pests, which seriously affect their yield and quality. , The breeding of disease-resistant varieties has always been a difficult problem. At present, the genetic transformation and in vitro mutation breeding of resistance genes have received extensive attention at home and abroad. Whether gene transformation and in vitro mutagenesis can be successfully applied to peanut breeding depends critically on the establishment of an efficient plant regeneration system in peanut tissue cultur...
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