Breeding technique of obtaining triploid grape and ploidy early identification using embryo

Abstract

The invention discloses a culture method for triploid grape culture by embryo rescue and early identification of ploidy. The invention aims to cultivate triploid grape by embryo rescue and overcome the problem of seed abortion of hybrid embryo between diploid and tetraploid grape cultivars in conventional breeding. After a hybrid seed is obtained, chromosome count is performed by the combination of flow cytometry and wall degradation hypotonic method to identify the ploidy of the hybrid progeny. The invention increases the breeding efficiency of triploid grape, accelerates the breed selection of triploid grape variety and provides a new approach to the culture of seedless grape variety.

Description

technical field [0001] The invention relates to plant breeding technology, in particular to a high-efficiency breeding technology for obtaining triploid grape hybrid seedlings by using embryo rescue culture, and carrying out systematic and comprehensive early identification of ploidy for the obtained hybrid offspring, and belongs to the field of biotechnology. Background technique [0002] Polyploidy breeding is an important way to cultivate new varieties of fruit trees. Its advantages lie in the characteristics of vigorous growth, thick branches, thick leaves, large fruits, high yield and strong adaptability. Budding, artificial chemical mutagenesis, artificial sexual hybridization, endosperm culture and anther culture, etc. Among them, the hybridization between diploid varieties and tetraploids is the most widely used, and results have been obtained in watermelons, bananas, and oranges, and triploid seedless fruits have been obtained. Grape is the second largest fruit in ...

AI Technical Summary

Problems solved by technology

[0004] Since the last century, through ovule culture, Japanese scholar Hiroyuki Yamashita has obtained triploid plants by crossing tetraploids and diploids of seeded grapes ("Journal of Horticultural Society" 1993, 62 (2): 249-255 ), then Li Shicheng ("Journal of Shanghai Agricultural Science" 1998, 14(4): 13-17), Pan Chunyun ("Journal of Shandong Agricultural University" 1998, 29(3): 299-302), Xu Haiying ("Journal of Fruit Tree" 2001 , 18(6): 317-320), Guo Yinshan ("Journal of Shenyang Agricultural University" 2005, 36(5): 606-608) and others also used embryo rescue technology to overcome the conventional breeding of grape diploid and tetraploid. Due to the obstacles of early abortion of hybrid embryos and low breeding efficiency, triploid hybrid seedlings have been successfully bred, but there is seldom a comprehensive ploidy test for the resulting hybrid seedlings, and a complete and efficient breeding system is lacking.

[0005] Usually polyploid identification methods include plant trait identification, cytological identification, molecular biological identification and other methods, while grape polyploidy is mostly identified by the number of chromosomes under a microscope, and it is also determined by flow cytometry. The chromosome counting method is intuitive and effective , but the material is limited to the shoot tip and root tip, and the operation is complicated

Flow cytometry is used to identify the DNA content of the plant to be tested. It can detect tens of thousands of cells in a short period of time, and can ensure the population characteristics of biological cells. The measurement is fast and the result is reliable, but it can only provide semi-quantitative measurement. result