Efficient and rapid sugarcane genetically modified method

Abstract

The invention belongs to the field of plant biotechnology, in particular to an efficient and rapid sugarcane genetically modified method, wherein spire tissues of sugarcane are taken as explants to induce embryonic callus; embryonic callus is taken as a receptor material infected by agrobacterium, and cultivates into embryoid, budlets and seedlings through differentiation, then resistance transformation is carried out to obtain the resistance transformation plants; then the DNA and RNA of resistance plants are extracted to carry out PCR and RT-PCR detection; the plants which are detected to be positive through PCR and RT-PCR detection are transplanted to a high tunnel, and then other molecular and physiology and biochemistry detections are carried out after the plants grow up. The invention can greatly improve transformation efficiency, shorten transformation time, obtain the transgenic plants which are detected to be positive by PCR and RT-PCR detections and only needs 4 months, and save transformation costs; and transplantation can be carried out after PCR and RT-PCR detections, thus reducing transplantation and management of a plurality of false resistance seedlings and lowering experimental cost.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and relates to a sugarcane transgenic breeding method, in particular to an efficient and rapid sugarcane genetically modified method. technical background [0002] Sugarcane is the most important sugar economic crop in the world, and it is also an important sugar economic crop in tropical and subtropical regions of my country. At present, my country is the third largest sugar producing country in the world. In the 2008 / 2009 crush season, the total sugar production in the country reached 12.43 million tons, of which cane sugar accounted for 94% of the total sugar production. In recent years, sugarcane has also developed into an important energy crop and an important raw material for the production of green fuel-ethanol. [0003] Since the modern sugarcane cultivars are interspecific hybrids of tropical species of Saccharum genus, Moshoumi and Indian species, during the long-term noble breeding ...

AI Technical Summary

Problems solved by technology

In the transformation process of Agrobacterium-mediated method, although the correct use of acetosyringone has greatly improved the transformation efficiency of Agrobacterium-mediated method, its transformation efficiency is about 3%, which still cannot meet the industrial development needs of transgenic engineering. Moreover, the Agrobacterium-mediated method still has deficiencies in the genetic transformation of sugarcane: (1) The adoption of sugarcane callus has always followed the callus subculture induction method of the early gene gun transformation method, and the differentiation efficiency of the callus itself is low. directly affects the final transformation efficiency; (2) after the co-cultivation of Agrobacterium and callus, it is difficult to completely remove the Agrobacterium, resulting in serious pollution during differentiation and regeneration; Screening is carried out at the stage of wound tissue differentiation, but because the shoots differentiated from sugarcane callus are small, it is difficult for the resistant shoots to grow taller, making it difficult to distinguish the resistant shoots from the shoots killed by the screening agent during the medium replacement process to open

(4) The conversion time is longer, more than 6 months