Co-Transformation Of Multiple Genes Into Wheat By Particle Bombardment

Wheat is one of the most important food crops in the world, and it is also the main crops in China. Meanwhile, Wheat、Rice and Maize are major grain crops in China, which has made a great contribution in the food supply. In addition, Wheat Scab(Fusarium head blight, FHB)caused by Fusarium graminearum, is one of the devastating disease of wheat in the cereal growing areas with warm and humid around the world. It can infect spike and stalk, but mainly infect the spike. It not only causes quality and yield loss, but also produces a variety of mycotoxins which threats to human and livestocks health. Wheat is hexaploid and has a large genome estimated at 16 Gb which comprises transposable elements and the most of the genome is repeated sequences, making traditional breeding very difficult. Furthermore, the resource to FHB and high production is also finite. Thus, it is important to use genetic engineering for high yield and FHB resistance to guarantee food safety.In this study, minimal cassettes of glycolate catabolic pathway-related genes and fingycin genes were introduced into the immature embryo callus of wheat variety Xiangmai 76 by particle bombardment. After Bar/Bialaphos or PMI/mannose screening, several positive plants were obtained. The main results were as follows:1. The co-transformation of glycolate catabolic pathway related genesWe transformed the minimal gene cassette At GDH, GCL and TSR and Bar from our lab into callus by particle bombardment, and get 3 positive plants. But one individual plant did not reach the heading stage. For PCR and RT-PCR of offspring, there are 3 aim genes in the 2 positive plants. We confer that the 2 plants can be inherited stably.2. The co-transformation of fengycin gene(1) With Bar selection gene, the minimal gene cassette FENB、FENE and Bar were transformed into wheat immature embryo callus. After Bialaphos screening and indentification, we get 2 positive plants. Through multiple generation in the greenhouse, the molecular identification shows that the genetic plants can be inherited stably.(2) With BAR selection gene, the minimal gene cassette FENC、SFP、Yc ZE and Bar were transformed into wheat immature embryo callus. After Bialaphos screening and indentification, we get 1 positive plant which contains SFP and Yc ZE. Though generation, the indentification of PCR indicate that the aim genes are transformed to wheat genome, and it can be inherited stably.(3) With Bar selection gene, the minimal gene cassette FENB、FENC and Bar were transformed into wheat immature embryo callus. After Bialaphos screening and indentification, we get 6 positive plants. Though greenhouse generation, the plants are T2 generation.(4) With PMI selection gene, the minimal gene cassettes FENB、FENE and PMI were transformed into wheat immature embryo callus. After mannose screening and indentification, we get 3 positive plants in T0 generation. Though generation, the indentification of PCR indicates that the 3 strains can be inherited stably.(5) With PMI selection gene, the minimal gene cassettes FENC、SFP、Yc ZE and PMI were transformed into wheat immature embryo callus. After mannose screening and indentification, we get 4 positive plants in T0 generation. Though greenhouse generation, the plants are T1 generation.