| Brown planthopper(Nilaparvata lugens St?l)(Hemiptera: Delphacidae) is the most serious pest on the rice, not only damage to the rice plants directly by sucking excess plant sap from the phloem but also indirectly by transmitting rice plant diseases of economic important. Currently, planting rich contain resistance genes is mainly way to fight the brown planthopper, but the brown planthopper can quickly adaption and overcome the resistance of rice continue to harm the rice by evolved a new biotype, which came from the brown planthopper and rice resistance gene coevolution. Brown planthopper prevention is a headache problem. Using molecular genetics theory and techniques can reveal the virulence of the brown planthopper use of molecular genetics theory and techniques can reveal the virulence use of molecular genetics theory and techniques can reveal the virulence of the brown planthopper genetic variation of the brown planthopper. In this study, used two different biotypes of brown planthopper to build mapping population, then combined SRAP marker, which widely used in the field of botany, to constructe genetic linkage map of brown planthopper, and evaluated the wild populations of brown planthopper virulence by four virulence phenotypes. Besides, developed whitebacked planthopper,(Sogatella furcifera)(Horváth)(Hemiptera: Delphacidae), genomic SSR markers to fill gaps in this field. The results are as follows:1.Identified three virulence phenotypes of inbred biotype 1 and biotype 2: The host preferences for Biotype 1 and Biotype 2 showed biotype 1 sensibility prefer rice TN1, while biotype 2 was no obvious preference for TN1 and Mudgo; survival rates for Biotype 1 and Biotype 2 on the TN1 and Mudgo rice plant showed survival rates of biotype 1 on the rice plant TN1 significant higher Mudgo, while biotype 2 was not significantly different on the two types of rice; growth rates for Biotype 1 and Biotype 2 on the TN1 and Mudgo rice plant showed females of 72 h weighing biotype 1(0.88) growth rates higher than biotype 2(0.91) on TN1 and no significantly different(P = 0.669), while growth rates of biotype 1(0.8) lower than biotype 2(0.95) on Mudgo, and have a very significant difference(P = 0.001). Phenotypic differences indicated two different biotypes of brown planthopper have significant differences in the genomic level, and they were suitable for constructing mapping population.2.By the orthogonal experimental design L16(45), five factors including Mg2 +, dNTPs, Taq DNA polymerase, primer concentration and DNA template concentrations amount in Nilaparvata lugens SRAP-PCR reaction system were optimized, and the optimal reaction system of the brown planthopper SRAP-PCR was established. The results show that: a suitable SRAP-PCR reacton system was constructed with the 10 ?L volumes containing 25 ng of template DNA, 0.3 ?mol/L of each of the two primers, 150 ?mol/L deoxynucleotide triphosphates(dNTPs), 3 mmol/L MgCl2, 1 × PCR buffer, and 2 unit of Taq DNA polymerase. We used the female of biotypes 1 and the male of biotype 2 monomer hybridization population to the optimal reaction system of the brown planthopper SRAP-PCR for authentication. The result showed that the bands were clear, with rich polymorphism map and co-dominant bands were found. It showed that the reaction system is stable and reliable.3.The maping population constructed by 14 generations of inbreeding biotype 1 male and 7 generations of inbreeding biotype 2 female hybridized F2 population, 92 females. The using 98 polymorphic SRAP markers to analye genotypes in this population, using JionMap 4 and MapChart 2.2 linkage analysis and draw a linkage map. Finally 14 linkage groups were constructed, including 188 sites, with a total length of 600.6 cM, the average interval 3.12 cM, 81.9% coverage map.4.In order to study the different biotypes of brown planthopper clustering, using 98 SRAP markers from genetic linkage map and five kinds of rice biotypes and a Leersia biotype, two closely related species of the brown planthopper, N. muiri China and N. bakeri(Muir). Received a total of 600 polymorphic bands, average 6.1 to each marker. Divided into two groups in the range of 0.5 to 0.82 of clustering, In the first group of five kinds of organisms on rice together, while brown planthopper on the Leersia was in a separate group. In the five biotype of rice brown planthopper, biotype 1 and biotype 3 was together, while biotype 2, biotype Y and biotype P was together, and biotypes Y and P was closely than biotype 2, in a single into a group. The results of clustering related to virulence of brown planthopper, suggesting that these polymorphic markers may be associated with virulence of brown planthopper.5.Using natural population brown planthopper fed on near isogenic lines rice 9311,which contained the resistance gene Bph9. Finally, 152 females brown planthopper to teste four virulence phenotype of brown planthopper, honeydew weight, weight gain, honeydew pH and abdominal development. Adult female was classified as avirulent or virulent based on the quantity of honeydew with the threshold of 10 mg/48 h, or weight gain with the threshold of 1 mg/48 h, or pH of honeydew with the threshold of 8 and abdomen phenotype was convex. honeydew weight or weight gain can be identified 30 virulence individuals and 27 individuals met the two indicators is 27, up to 90% of the total. Correlation analysis of honeydew and weight gain derived, r = 0.808(P<0.01), is highly relevant; pH of honeydew identified 2 avirulence individuals and 20 virulence individuals, 2 avirulence individuals correspond to honeydew weight and weight gain phenotype, but the virulence of 20 individuals matched with honeydew weight and weight gain phenotype were 60% and 65%; abdominal development were identified 32 individuals were virulence,and matched with honeydew, weight gain and honeydew pH consistent phenotype were 84.3%, 90.6% and 100%. This study suggests that only both weight gain and honeydew weight exceed 1 ? and 10 ? individual to determine for virulence individual.6.The whitebacked planthopper,Sogatella furcifera(Horváth)(Hemiptera: Delphacidae), is one of the most harmful pests of rice. In this study, 18 polymorphic microsatellite markers were developed from S. furcifera genomic libraries using the fast isolation by amplified fragment length polymorphism of sequences containing repeats method. Microsatellite polymorphism was investigated using 32 individuals from one natural population. These 18 simple sequence repeat markers showed a number of alleles that ranged from 3 to 15 and had observed and expected heterozygosities that ranged from 0.094 to 0.871 and from0.148 to 0.924, respectively. The high cross-species transferability of these markers was evaluated in three other planthopper species: Nilaparvata lugens, N. muiri China, and N. bakeri(Muir). These microsatellite markers will provide powerful tools for population genetic and ecological studies of this pest and its related species in the future. |
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