Study On Male Sterility Of Winter Wheat Induced By A New Chemical Male Sterility Agent CH1

Wheat(Triticum aestivum L.)is the most important cereal crop in China,and its yield improvement is the key to ensure food security.Selection and breeding of hybrid wheat varieties to exploit their hybrid advantage and thus increase the yield of wheat is a proven way.At this stage,in the utilization of wheat hybrid advantage,induction treatment of male sterility using chemical androgenicides can be used for breeding hybrid wheat based on conventional breeding of wheat.It provides a new idea for the selection and breeding of common wheat varieties.However,among the widely used androgenic agents for wheat,all of them have one thing in common,that is,they are all special wheat androgenic agents,and the process of their production is not only very complicated,but also the production cost is high.Therefore,it is of practical significance to screen out new chemical androgenicides that are inexpensive,easy to produce and highly efficient,so as to reduce the cost of hybrid wheat production and to promote the use of hybrid advantage in wheat.In order to elucidate the effect of the new chemical androgenic agent CH1 in inducing male sterility in wheat and its sterility mechanism,and to provide theoretical and technical support for the development and application of chemical androgenic agents in wheat.In this study,the new chemical androgenicide CH1 was used in wheat varieties Pu Bing 151,Bainong 607 and Bainong 8822,and six dose levels(0,10,15,20,25 and 30 g/hm~2)and seven surface activators(NE1805,NE1808,NE1815,NE1820,AC1810,AC1820,AC1220)for CH1 dose and surface activator screening,and three 40%vinblastine levels(0,5,10 m L/L)at 15 g/hm~2dose level for CH1-vinblastine interaction test.The RNA of anthers of chemically killed sterile lines and control normal fertile lines were also sequenced and analyzed using transcriptome sequencing technology,and the genes with their differential expression ploidy were obtained from the sequencing data,while the screened genes were validated using real-time quantitative PCR technology.The main findings obtained in this thesis are as follows:1.Using Pubing 151,Bainong 607 and Bainong 8822 as test materials,when wheat was sprayed with different doses of CH1 at Feekes 8.0 during the growth period,CH1 was able to induce sterility of Pubing 151 and Bainong 8822 at a dose of 10 g/hm~2to produce more than 97%,while the sterility rate of Bainong 607 was lower(62.2%);however,when CH1 was treated at doses of 15～30 g/hm~2dose treatment,the sterility rates of all three genotypes of wheat varieties were above 98%,indicating that CH1 is broad-spectrum in inducing male sterility in wheat.2.Using Pubing 151 as the test material,CH1 was selected at a dose concentration of10 mg/hm~2,and the screening test of seven surface activators showed that the addition of different surface activators had different degrees of response to the plant flag leaf,among which the addition of NE1805,AC1810 and AC1220 caused more severe damage to the plant flag leaf,all of which exhibited dry leaf tips and partial area loss of green,while the others only All surface activators had no significant effect on wheat spike development,but reduced plant height,with the highest reduction(45.3 cm)after the addition of NE1805;investigating the sterility rate,it was found that under the chemical androgenic agent CH110 g/hm~2dose,the sterility rate was 24.3%for NE1805 and 82.0%for AC1220,except for the other surface activators.Except for NE1805 with a sterility rate of 24.3%and AC1220with a sterility rate of 82.0%,several other surface activators were able to induce sterility rates of more than 96%;to investigate the fertility rate of artificial pollination,the fertility rates were higher after adding surface activator NE1808,NE1815 and NE1820 treatments,54.0%,60.3%and 57.0%,respectively,while the fertility rates after other surface activator treatments were lower.The results indicated that the new chemical androgenic agent CH1had better androgenic effect when used with the surface activator NE1820.3.The intercropping test of CH1 and vinblastine was conducted at 15 g/hm~2dose level of CH1 and three levels(0,5 and 10 m L/L)of 40%vinblastine with Pubing 151,Bainong607 and Bainong 8822 as test materials.The results showed that the reaction of the flag leaves of the three genotypes to the addition of vinyl chloride was significantly reduced,and the degree of dry leaf tips became smaller;as the concentration of vinyl chloride increased,the plant height of the three genotypes gradually decreased and the tassel stage was pushed back,but there was no significant change in spike length.However,after the addition of 10 m L/L of ethylene glycol,the sterility rates of Pubing 151 and Bainong 607both showed a certain degree of decrease,97.3%and 91.9%,respectively.Investigating the pollination set rate,it was found that the addition of 5 m L/L ethylene glycol treatment increased the pollination set rate of the three genotypes,but the pollination set rate of the three genotypes decreased significantly after the addition of 10 m L/L ethylene glycol.This indicates that the addition of 5 ml/L of ethylene glycol at low concentrations can reduce the drug damage,reduce the plant height,and promote the increase of pollination rate,which is beneficial to seed production.4.The above study showed that when the chemical androgenic agent CH1 was added at 15 g/hm~2with 1%NE1820 and 5 m L/L of 40%vinblastine could produce complete male sterility in wheat,reduce plant height,and obtain high pollination rate.It can be used as a candidate for hybrid wheat seed production,and further research and utilization.5.Transcriptomic analysis of anther RNA of chemo-killed sterile lines and control normal fertile lines was performed using transcriptome sequencing technology with wheat variety Pubing 151 as test material.The results showed that there were 3895 differentially expressed genes,including 1334 up-regulated genes and 2561 down-regulated genes,in the chemo-killed sterile lines compared with the control normal fertile lines.The largest number of genes were annotated as hydrolase activity and protein binding,with 611 and464 genes,respectively,followed by ATP binding and carbohydrate metabolic process and carbohydrate metabolic process with 353 and 275 genes,respectively,followed by protein domain specific binding and the mitochondrial proton-transport ATP synthase complex pathway(mitochondrial proton-transporting ATP synthase complex had the lowest number of differential genes,both with three genes.6.The five genes screened from the transcriptome data with large expression difference ploidy were validated using real-time quantitative PCR.The results showed that the five genes showed different up-and down-regulation trends in the mononuclear,binuclear and trinuclear phases after CH1 treatment compared with the control,but all five genes showed a significant down-regulation trend at the binuclear phase,which was consistent with the transcriptome sequencing results,and the down-regulated expression of these five genes could possibly lead to the abortion of pollen grains.