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Genetic Diversity Analysis of Sogatella furcifera (Horváth) (Hemiptera: Delphacidae) by Using Novel Microsatellite Markers : 초위성체 마커를 이용한 흰등멸구 개체군의 유전적 다양성 분석

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dc.contributor.advisor이준호-
dc.contributor.author남화연-
dc.date.accessioned2017-07-14T06:43:01Z-
dc.date.available2017-07-14T06:43:01Z-
dc.date.issued2014-08-
dc.identifier.other000000021847-
dc.identifier.urihttps://hdl.handle.net/10371/125875-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 농생명공학부, 2014. 8. 이준호.-
dc.description.abstractWhite-backed planthopper, Sogatella furcifera (Horváth) (Hemiptera: Delphacidae), is known as a major rice and long-range migratory pest in Asia. Microsatellite marker (SSR) is widely used to perceive the origins and genetic diversity of insect pest. Samples were collected from Laos, Vietnam and 3 localities in Bangladesh, and we characterized novel 10 microsatellite loci of S. furcifera by using next-generation Roche 454 pyrosequencing technologies. We used 40 adult individuals collected from Shinan to test utility of ten microsatellite loci. The average of alleles per locus were 7.7. The mean of observed (HO) and expected heterozygosities (HE) were 0.648
and 0.748 respectively. These new microsatellite markers will be widely used in future ecological genetic studies of S. furcifera, including gene flow and genetic diversity among the population that are necessary for effective management and observing of the species.
Furthermore, we used these markers to determine genetic diversity for S. furcifera specimens collected from Korea (Shin-an), Laos, Nepal, Thailand, Vietnam and three different sites of Bangladesh in 2012, and Bangladesh, China, Nepal ,Thailand, and fifteen different sites of Korea in 2013. The genetic variability estimates for each S. furcifera population infer from the ten microsatellite loci included the observed (HO) and expected (HE) heterozygosity and the inbreeding coefficient. HO ranged from 0.259 – 0.604 (mean = 0.435) and HE ranged from 0.488 - 0.768 (mean = 0.669). The FIS ranged from 0.1867 in WBPH_T17 to 0.5855 in WBPH_T11, with mean of 0.3598 across loci. In genetic variability estimates for each S. furcifera population in Asia, inferred from ten microsatellite markers, Shinan has a highest number of HO 0.648. This result shows the possibility of variation of the migration source. We speculate Laos, Vietnam and China as the origin of S. furcifera in Korea by the low number of FIS and high number of HO. The genetic variability estimates for each S. furcifera population in Korea, Changnyung, Gosung, Milyang and Shinan show high estimation of HO. All of these sites located in south and southeast part of Korea and may assume
variability of migration origin. This study provides useful data to forecast the migration and origin of S. furcifera. This information about migration pattern may develop sustainable pest management strategies of this long-range migratory pest.		-
dc.description.tableofcontentsList of Contents

Abstract ------------------------------------------------------------------------------------- I List of contents ------------------------------------------------------------------------- IV
List of Tables ----------------------------------------------------------------------------- VI
List of Figures -------------------------------------------------------------------------- VIII
I. Literature Review ---------------------------------------------------------------------- 1
1-1. General characteristic of white backed planthopper ------------------ 2
1-2. New generation sequencing (NGS) --------------------------------------- 3
1-3. Microsatellite marker---------------------------------------------------------- 6
III. Developing Novel microsatellite markers of Sogatella furcifera (Horváth)
(Hemiptera: Delphacidae)---------------------------------------------------------- 7
2-1. Introduction ---------------------------------------------------------------------- 8
2-2. Materials and Methods ------------------------------------------------------ 10
2-2-1. Microsatellite marker development ------------------------------ 10
2-2-2. Roche 454 library preparation and sequencing -------------- 13
2-2-3. Microsatellite marker development ------------------------------ 16
2-2-4. DNA sample and Microsatellite genotyping ------------------- 17
2-2-5. Data analysis ---------------------------------------------------------- 17
2-3. Results -------------------------------------------------------------------------- 18
2-4. Discussion --------------------------------------------------------------------- 25
III. Genetic diversity analysis of Sogatella furcifera (Horváth) (Hemiptera:
Delphacidae) in Asia -------------------------------------------------------------- 27
3-1. Introduction -------------------------------------------------------------------- 28
3-2. Material and Methods ------------------------------------------------------- 30
3-2-1. Study insect and sample collection ------------------------------ 30
3-2-2. Microsatellite genotyping ------------------------------------------- 35
3-2-3. Statistical analysis --------------------------------------------------- 36
3-3. Results -------------------------------------------------------------------------- 37
3-4. Discussion --------------------------------------------------------------------- 44
VI. Literature Cited --------------------------------------------------------------------- 45
초록 --------------------------------------------------------------------------------------- 53

List of Tables

Table 1. Sampling information of Sogatella furcifera ----------------------------11

Table 2. Primer sequences used to create pooled plant hopper libraries for sequencing on the Roche 454. Underlined, grey highlighted and black highlighted regions respectively correspond to the key, Midtag (4 or 11), and adapter sequences --------------------------------------------------------------------- 15

Table 3. Summary of newly designed primers ---------------------------------- 20

Table 4. Modified twelve proper microsatellite markers --------------------- 23

Table 5. Characteristics of the ten S. furcifera microsatellite loci tested in S. furcifera specimens from countries ------------------------------------------------- 24

Table 6. 2012 and 2013 samples of S. furcifera --------------------------------- 33

Table 7. Characteristics of the ten S. furcifera microsatellite loci tested in S. furcifera specimens from countries ------------------------------------------------- 39

Table 8. Genetic variability estimates for each S. furcifera population in Asia, deduced from ten microsatellite loci. Number of alleles, expected heterozygosity (HE), observed heterozygosity (HO) at HWE and inbreeding coefficient (FIS) -------------------------------------------------------------------------- 40

Table 9. Genetic variability estimates for each S. furcifera population in Korea, deduced from ten microsatellite loci. Number of alleles, expected heterozygosity (HE), observed heterozygosity (HO) at HWE and inbreeding coefficient (FIS) -------------------------------------------------------------------------- 42

List of Figures

Fig 1. Map of S. furcifera collecting sites ----------------------------------------- 12

Fig 2. Selecting new microsatellite markers by using GeneMapper program ----------------------------------------------------------------------------------------------- 22

Fig 3. Sampling sites in East Asia (2012) ----------------------------------------- 31

Fig 4. Sampling sites in East Asia (2013) ----------------------------------------- 31

Figure 5. Map of S. furcifera collecting sites in Korea (2013) ---------------- 32		-
dc.formatapplication/pdf-
dc.format.extent928336 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoko-
dc.publisher서울대학교 대학원-
dc.subjectwhite backed planthopper-
dc.subjectSogatella furcifera-
dc.subjectgenetic diversity-
dc.subjectmicrosatellite marker-
dc.subjectnext generation sequencing-
dc.subject.ddc630-
dc.titleGenetic Diversity Analysis of Sogatella furcifera (Horváth) (Hemiptera: Delphacidae) by Using Novel Microsatellite Markers-
dc.title.alternative초위성체 마커를 이용한 흰등멸구 개체군의 유전적 다양성 분석-
dc.typeThesis-
dc.description.degreeMaster-
dc.citation.pagesVIII, 56-
dc.contributor.affiliation농업생명과학대학 농생명공학부-
dc.date.awarded2014-08-
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