Breeding of anti-ear sprouting transgenic wheat

The Henan Provincial Outstanding Talent Innovation Fund project, funded by the National Wheat Engineering Technology Research Center Dr. Yin Yin, “Cultivation of Anti-Frozen Germ Wheat”, using modern genetic engineering technology, developed a transgenic technology that inhibits mature wheat germinating on a continuous cloudy and rainy day. The bottleneck technical problems that affect wheat yield and quality were solved, and a new line of transgenic wheat resistant to ear sprouting was developed. At present, the project has completed a pilot test in Henan Province, participated in the national safety evaluation, and was included in the National Agricultural Science and Technology Achievement Transformation Project. After the project is promoted, it will produce greater disaster reduction and social benefits. The new technology of GM wheat has entered the pilot stage at Henan Agricultural University, a wheat genetically modified technology aimed at inhibiting the germination of mature wheat on a continuous rainy day. In mid-June 2004, a pilot test was conducted at Henan Agricultural University to use this technology to cultivate transgenic plants. The wheat germ-resistant new strains were also tested in Henan Province at the same time. Experts believe that the successful development and application of wheat anti-ear sprouting technology will play an active role in the safe production of wheat in China. According to reports, the phenomenon of ear sprouting during the rain before wheat harvest is an important issue in wheat production in major wheat producing countries in the world. Due to the impact of ear sprouting, the world wheat output has lost nearly 100 million tons annually. In the Huanghuai wheat district of China, many large-scale wheat sprouts occurred due to the rain during the ripening period, which in turn caused the broken seeds and the inability of the flour to be eaten. In the northern wheat region, although there was no rainy season, individual wheat harvest periods also occurred. Continuous rain caused wheat germination. As the sprouting of the ear will lead to a series of physiological and biochemical changes in the wheat seeds, protein, starch and other storage substances will be decomposed, resulting in a decrease in grain yield, especially the nutritional quality and processing quality of the flour. The loss of wheat seed value will cause wheat production. Serious economic loss. Therefore, wheat germination is a problem that has been widely concerned by wheat breeders and physiologists at home and abroad for a long time. At present, most wheat cultivars used in production have certain ear sprouts. Yin Hao, the deputy director of the National Wheat Engineering Technology Research Center of Henan Agricultural University, introduced that studies on the biological function of thioredoxin Trx and the antisense gene mechanism at home and abroad have provided a new approach for the establishment of wheat germplasm resources resistant to ear sprouting. Trx is a functional protein that catalyzes the oxidation and reduction of disulfide bonds. It can change the activity of important enzymes such as amylase and protein enzymes in seeds, increase the water solubility of storage proteins and promote the hydrolysis of nutrients, and germinate the seeds of crops. The supply of nutrients during the process has an important regulatory role. The trxs gene and Trx gene from the blue-duck grass belong to the thioredoxin gene family, and their protein expression products have similar biological activities. The introduction of the antisense trxs gene into wheat is expected to reduce the activity of Trx in seeds, inhibit the activity of amylase and protein enzymes, slow the hydrolysis of starch, protein and carbon and nitrogen metabolism during seed germination, and ultimately lead to the weakening of germination ability of transgenic wheat seeds. The research group began to carry out the research on the introduction of anti-trxs gene into wheat from 1997. Under the support of the national “863” project, the national project for research on transgenic plants and industrialization, and the Henan Outstanding Talent Innovation Fund, the research adopted the transformation of the gene gun. In the technology, genetic transformation was carried out on more than ten varieties and tens of thousands of immature embryos in the Huanghuai wheat region. Through the induction, selection and plant regeneration of callus from immature embryos, transgenic regenerated plants were obtained and the transgenic wheat offspring strains were obtained. Department of tracking molecular detection and homozygous line screening, has now been obtained in the wheat genome genetically stable gene of the fourth generation of wheat. Through the molecular detection of the target gene, the genetic stability of the target gene in the wheat genome was determined; through the determination of biochemical physiological characteristics of the transgenic wheat grain and endosperm, it was proved that the target gene can be normally expressed in some strains of transgenic wheat; The transgenic plants with stable genetic inheritance and expression were subjected to a simulated rainfall-precipitation germination test, confirming that some of the transgenic lines had strong resistance to ear sprouting compared to the control group. According to Yin Hao, they have obtained four ability to resist germination based on their integration in the genome, molecular testing results, physiological and biochemical changes of transgenic wheat grains, and performance of the transgenic lines in the anti-ear sprouting test. Transgenic homozygous lines, and the results showed that the antisense trxs gene has been integrated into the wheat genome and can be stably inherited. The introduction and expression of the antisense trxs gene directly interferes with the expression of the wheat's own Trx gene. The level of thioredoxin in the transgenic wheat grain decreased. By inhibiting the thioredoxin activity of wheat itself, the activities of high amperage amylase isoenzymes are inhibited, and the level and solubility of free sulfhydryl groups in stored proteins are reduced, affecting the hydrolysis and mobilization of the stored material of the seeds and normal carbon and nitrogen metabolism. Therefore, the main reason for the anti-germination ability of genetically modified wheat is that the nutrients required for germination cannot be effectively supplied.

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