Genetic transformation method using sweet sorghum young ear or young ear induced callus as explant

A genetic transformation method and callus technology, applied in the field of genetic transformation to achieve the effect of high callus induction rate

Inactive Publication Date: 2011-01-19
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Genetic transformation method using sweet sorghum young ear or young ear induced callus as explant
  • Genetic transformation method using sweet sorghum young ear or young ear induced callus as explant
  • Genetic transformation method using sweet sorghum young ear or young ear induced callus as explant

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Embodiment 1

[0034] Example 1 Establishment of Tissue Culture Regeneration System for Young Ears of Sweet Sorghum

[0035] 1.1 Selection and preparation of ear materials for sweet sorghum tissue culture

[0036] The present invention gets the young ear of about 2~5cm in the flag leaf stage, carries out surface disinfection with 70% alcohol for 3min, rinses with sterile water for several times; in the ultra-clean bench, cut into slices of uniform size with a knife and inoculate them on the induction medium [MS Basal medium+2,4-D 2mg / L+ZT (zeatin) 2.0mg / L+PVP (polyvinylpyrrolidone) 10mg / L+ascorbic acid 10mg / L+hydrolyzed casein 500mg / L+sucrose 30g / L+agar 7.2g / L, pH 5.8] placed at 20-21°C for 3-4 weeks in dark culture to start dedifferentiation and callus formation; then select callus with rapid growth, crisp texture and bright color for subculture [induction culture Base + KT (kinetin) 0.2mg / L], subculture every 7 to 10 days. Select the callus with good growth state and connect it to the d...

Embodiment 2

[0053] Embodiment 2. Agrobacterium transformation

[0054] 2.1 Media used for Agrobacterium transformation:

[0055] Agrobacterium pre-medium: 1 / 2MS inorganic salt + yeast extract 2.5g / L + sucrose 30g / L + glucose 60g / L + AS100μmol / L + 125mM streptomycin (SM) + 100mM kanamycin (Kan), pH 5.6;

[0056] Agrobacterium suspension: 1 / 2MS inorganic salt+yeast extract 2.5g / L+2,4-D 1.5mg / L+sucrose 68.5g / L+glucose 36g / L+AS 100μmol / L, pH 5.2.

[0057] Pre-medium YS-P: MS inorganic salt+MS organic matter+2,4-D 2mg / L+ZT 2mg / L+KT 0.2mg / L+hydrolyzed casein 500mg / L+PVP 10mg / L+AS 100μmol / L+sucrose 30g / L+agar 8g / L, pH5.8;

[0058] Co-culture medium YS-C: MS inorganic salt + MS organic matter + 2,4-D 2mg / L + ZT 2mg / L + KT 0.2mg / L + hydrolyzed casein 500mg / L + ascorbic acid 10mg / L + PVP 500mg / L + AS 100μmol / L+sucrose 30g / L+agar 8g / L, pH5.6;

[0059] Screening medium YS-S: MS inorganic salt+MS organic matter+2,4-D 2mg / L+ZT 2mg / L+KT 0.2mg / L+hydrolyzed casein 500mg / L+ascorbic acid 10mg / L+PVP 10m...

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Abstract

The invention relates to a genetic transformation method using sweet sorghum young ear or young ear induced callus as an explant, belonging to the technical field of plant genetic engineering. In the invention, the agrobacterium rhizogenes meditation method is adopted to transform foreign genes for the sweet sorghum young ear explant induced callus; resistant healing is obtained by carrying out preculture, coculture and screening culture in sequence on the sweet sorghum young ear explant induced callus; and then the transformed plant is obtained by carrying out successive differentiation culture, budding, rooting and transplanting on the sweet sorghum young ear explant induced callus. In addition, the established genetic transformation system is used to transform the sweet sorghum plant to obtain the genetically modified pest-resistant plant. The method of the invention provides technical support for researches on using the biotechnology to improve quality of the sweet sorghum and increase the stress resistance and the like and lays very good foundation for accelerating the basic research on the sweet sorghum.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, in particular to a genetic transformation method using sweet sorghum young ears or callus induced by young ears as explants. Background technique [0002] Sweet sorghum is a variant of common sorghum. Its growth water consumption is only 1 / 3 of that of corn, and its yield per mu can be as high as 5-10 tons. Its stalk juice is an ideal biomass raw material for fermentation to produce alcohol and yeast. The new technology of using sweet sorghum instead of corn and other food crops as raw materials to produce alcohol can not only save food, reduce costs, effectively promote the fermentation industry, revitalize the alcohol production industry, but also promote the development of renewable energy and animal husbandry. The key to the development of sweet sorghum industry lies in the selection and efficient cultivation of new varieties of sweet sorghum. Traditional breeding technique...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 黄大昉朱莉郎志宏李桂英
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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