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Method for establishing efficient regeneration system for lagenaria siceraria by taking cotyledonary nodes as explants

A high-efficiency regeneration and explant technology, applied in the field of plant genetic engineering, can solve the problems of narrow genetic basis of cucurbit crops, unsuccessful application of cucurbits, poor repeatability, etc., and achieves the effects of maintaining genetic stability, easy implementation and simple operation.

Inactive Publication Date: 2016-07-20
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

Although other crops of Cucurbitaceae have established regeneration systems, due to the narrow genetic basis of Cucurbitaceae crops, the existing regeneration systems cannot be successfully applied to cucurbits, and the reproducibility is poor in the actual operation process, and the regeneration rate is extremely low

Method used

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  • Method for establishing efficient regeneration system for lagenaria siceraria by taking cotyledonary nodes as explants
  • Method for establishing efficient regeneration system for lagenaria siceraria by taking cotyledonary nodes as explants
  • Method for establishing efficient regeneration system for lagenaria siceraria by taking cotyledonary nodes as explants

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Embodiment Construction

[0033] 1. Cultivation of aseptic seedlings of the gourd rootstock variety Chaofeng Kangshengwang: pick the mature and plump seeds of the gourd rootstock Chaofeng Kangshengwang, manually remove the shells to obtain the seeds, and sterilize the seeds with 70% absolute ethanol for 40 seconds. Rinse with sterile water for 2-3 times, then disinfect with 10% NaClO for 8-12 minutes, rinse with sterile water for 2-3 times, place on sterile filter paper to control the moisture, and then transfer to hormone-free adventitious buds Induction medium MS1 (such as figure 1 shown), cultured in the dark at 25°C until the seeds germinated to obtain sterile seedlings (such as figure 2 shown), wherein adventitious bud induction medium MS1 includes: MS medium, 3.0 mg·L -1 6-BA, 0.01mg·L -1 2,4-D, 0.3mg·L -1 AgNO 3 , 30g·L -1 of sucrose and 8g·L -1 The agar has a pH of 5.6-6.0, preferably 5.8.

[0034] 2. Get the aseptic seedling 3d after the seed germination, cut off the embryo and hypocot...

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Abstract

The invention particularly relates to a method for establishing an efficient regeneration system for lagenaria siceraria by taking cotyledonary nodes as explants. The method includes steps: (1) selecting ripe and plump seeds, dehulling to obtain kernels, sterilizing, transferring to an MS basal culture medium, and culturing to obtain sterile seedlings; (2) taking the sterile seedlings to obtain cotyledonary node explants; (3) inoculating the cotyledonary node explants into an adventitious bud induction medium MS1, and culturing to obtain adventitious bud clusters; (4) stripping the adventitious bud clusters off the explants, transferring to a subculture medium MS2, and separating the adventitious bud clusters to obtain single adventitious buds when the adventitious buds grow to 3-4cm in height; (5) transferring the single adventitious buds to a rooting medium MS3, and culturing until rooting is realized; (6) subjecting the rooted regenerated plants to hardening treatment; (7) after hardening is finished, transplanting seedlings into a seedling culture substrate, and performing greenhouse culture. The method is high in bud induction rate, simple in operation, easy to implement and applicable to gene manipulation including germplasm innovation, genetic improvement and the like of lagenaria siceraria.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a method for establishing a high-efficiency regeneration system of gourd by taking cotyledon nodes as explants. Background technique [0002] Cucurbit (Lagenaria sisiceraria Standl.) belongs to Cucurbitaceae (Cucurbitaceae), and is an annual herbaceous plant. Originated in Africa. Its tender fruit is edible, and the mature fruit can be used as a container. In addition, gourds also have a special purpose, that is, they can be used as rootstocks for grafting watermelons and melons. The initial purpose of grafting gourds was to control watermelon wilt (Fusariumoxysporumf.sp.niveum). Fusarium wilt is a devastating soil-borne disease in watermelon production. It belongs to the watermelon specialization type of Fusarium oxysporum. It will occur throughout the growth period of watermelon. The pathogen mainly infects with mycelia, chlamydospores or sclero...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 张曼羊杏平徐锦华胡雪丹刘广姚协丰李苹芳任润生
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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