Induction of tung tree hypocotyls callus and method for efficiently regenerating plants

A callus induction and high-efficiency regeneration technology, applied in the fields of botanical equipment and methods, plant regeneration, horticultural methods, etc., can solve the transformation obstacles of the Aleuca vulgaris genetic system, inability to obtain transgenic plants, low kanamycin resistance, etc. problem, to achieve high vigor and differentiation and regeneration ability, not easy to die, good seedling effect

Active Publication Date: 2014-10-15
CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The research on the in vitro culture of tung tree is very late. Up to now, there are only reports of obtaining regenerated plants from the isolated leaves of tung tree through the organogenesis pathway. However, due to the tenderness of the leaves, they are particularly vulnerable to injury during Agrobacterium infection. It is a great obstacle to realize the transformation of the genetic system of tung tree, and the leaves are easy to lose water, the resistance to kanamycin is low, and they are easy to die during the selection process of kanamycin, while the hypocotyl is Embryogenic organs developed through zygotic embryos have strong embryogenicity, high vitality and ability to differentiate and regenerate, and have broad prospects in the transformation of plant genetic systems.
At present, transgenic plants obtained by infecting hypocotyls with Agrobacterium have been successful in rapeseed and citrus, and these plants can also obtain regenerated plants from detached leaves, but during the process of Agrobacterium infection, as well as in Kanna In the selection of kanamycin, the leaves are easy to die, and the transgenic plants cannot be obtained. On the contrary, the hypocotyls of these plants are relatively resistant to kanamycin, which is conducive to the mediation, transformation and regeneration of the target gene.

Method used

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  • Induction of tung tree hypocotyls callus and method for efficiently regenerating plants
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  • Induction of tung tree hypocotyls callus and method for efficiently regenerating plants

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Embodiment 1

[0045] Do the following in order:

[0046] 1. Select mature tung tree seeds of the same year, remove the seed coat, rinse the seed kernels with tap water for 5-8 minutes, then soak for 15-20 hours, change the water 3-4 times in the middle, and then use 0.1% HgCl 2 Disinfect for 3-5 minutes, rinse with sterile water 4-6 times, inoculate in 1 / 2 MS basic medium, culture in dark for 5 days, then switch to light and culture for 25 days, the hypocotyl height can reach 3-4cm, Cut into small pieces of 0.5-1 cm under sterile conditions, and inoculate them in WPM medium + 5.0-10.0 mg / L6-BA + 1.0 mg / L KT + 0.1 mg / L NAA to induce healing Injured tissue, cultured in the dark for 3 days, then cultivated in the light for 10 days, the highest induction rate can reach 100%. Additional sucrose 30g / L, agar 7g / L, pH5.5. The culture temperature is (26±1)°C, the light intensity is 2100-2200lx, and the light time is 12-14h / d; (see attached figure 2 )

[0047] Table 1 Effects of different hormo...

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Abstract

The invention discloses induction of tung tree hypocotyls callus and a method for efficiently regenerating plants, and belongs to the field of plant cell engineering. According to the invention, through the processes of inducing the callus by the hypocotyl of the tung oil tree, inducing an adventitious bud by the callus, performing transgenerational multiplication culture, performing strong seedling culture, performing rooting culture, performing acclimatization and transplantation and the like, a way for quickly breeding good tung oil tree plants is provided, and a firm foundation for later improving the resistance of the tung oil tree, accelerating the molecular breeding process of the tung oil tree, improving the oil quality and other properties, and establishing the genetic system of the tung oil tree in a genetic engineering method is built.

Description

technical field [0001] The invention belongs to the field of tung tree cell engineering, and in particular relates to a method for inducing and efficiently regenerating plants by using tung tree hypocotyl callus tissue. Background technique [0002] Tung tung is the general name of Euphorbiaceae (Euphorbiaceae) Tung genus (Vernicia) plants, is an important industrial oil tree species, and together with Camellia oleifera, walnut and Chinese tallow tree are called the four major woody oil tree species in my country. The oil extracted from the seeds of tung oil is called tung oil, which is the highest quality plant drying oil in the world, and is an excellent raw material for paints and printing inks. Tung oil has excellent characteristics such as fast drying, strong adhesion, light specific gravity, rich luster, acid and alkali resistance, high temperature resistance, frost crack resistance, and water resistance. In addition, tung oil is also an excellent coating and the basi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 谭晓风李泽张琳卢锟龙洪旭吴玲利林青吕佳斌
Owner CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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