Effects of Space-induced Mutation and Analysis of Polymorphsim by AFLP Molecular Marker in Sesame

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1 华北农学报 2013,28 ( 1 ) : AFLP D15 05N01 06ms198 5 SP3 SP4 11 AFLP 53 cm 77 cm 16 cm 122 / g kg /hm 2 36 cm 30 cm 19 cm 197 / g kg /hm 2 11 SP4 AFLP ~ % ~ ~ % ~ AFLP S A Effects of Space-induced Mutation and Analysis of Polymorphsim by AFLP Molecular Marker in Sesame GAO Tong-mei 1 WEI Shuang-ling 1 LI Chun-ming 1 ZHANG Hai-yang 1 LI Feng 1 HE Song-tao 2 1. Henan Sesame Research Centre Zhengzhou China 2. Henan Institute of Science and Technology Xinxiang China Abstract The seeds of five sesame varieties Yuzhi No. 11 Japan black sesame Zhengzhi D15 Zhengzhi 05N01 and Zhengzhi 06ms198 were carried to space for inducing mutation by Shijian 8 satellite. The agronomic trait and resistant variants of SP3 and SP4 were observed and the genetic diversity of mutation descendants of Yuzhi NO. 11 and Japan black sesame was analyzed using 16 pair of AFLP markers. The results showed that the variants of leaf capsule flower color childbearing period plant height first capsule height yellow apex length capsule number per plant 1000-seed weight yield and disease resistance were observed in SP3 and SP4 of different sesame varieties but the variation frequency and variation type were significantly different among different sesame varieties. The agronomic traits showed more negative mutations than positive mutations but resistance traits had more positive mutations. Compared to CK without space mutation the plant height first capsule height yellow apex length capsule number per plant 1000-seed weight and yield increased by 53 cm 77 cm 16 cm 122 /plant g kg /ha for positive mutation respectively and decreased by 36 cm 30 cm 19 cm 197 /plant g kg /ha for negative mutation respectively. AFLP analysis indicated that the genetic distance of Yuzhi No. 11 was of which 72% was from to and the average genetic distance was The genetic distance of Japan black sesame was from to of which 37. 5% was from to and the average was It was concluded that the space-induced mutation had no direction and great randomness the sen CARS

2 228 华北农学报 28 卷 sitivity was different in sesame varieties. Key words Sesame Space-induced AFLP molecular marker Analysis of polymorphsim SP CK SP2 50% 50% 2009 SP2 SP3 3-4 SP1 SP2 SP SP4 RAPD 3 5 DNA 7 = - / 100% AFLP DNA SP2 SP3-80 AFLP DNA CTAB DNA 1% DNA ND DNA 5 DNA 100 ng /μl AFLP 9 AFLP EcoRⅠ EA 5'-CTCGTAGACTGCGTACC- 3' 3'-CTGACGCATGGTTAA-5' D15 05N01 06ms min SPn CK D15 05N01 06ms198 SP3 Mse Ⅰ MA 5'-GACGATGAGTCCTGAG- 3' 3'-TACTCAGGACTCAT-5' DNA 100 ng /μl 2. 5 μl EcoR Ⅰ 10 U /μl 0. 5 μl MseⅠ 10 U /μl Ⅰ0. 5 μl 10 Buffer 1. 5 μl ddh 2 O 10 μl 15 μl 37 3 h 65 3 h μl EcoR Ⅰ 5

3 1 期高桐梅等 : 芝麻太空诱变效应及 AFLP 标记检测 229 μmol 2. 0 μl MseⅠ 50 μmol 2. 0 μl T4 DNA 53 ~ - 36 cm 77 ~ - 30 cm 3 U /μl 0. 4 μl 10 Buffer 1. 0 μl 16 ~ - 19 cm 122 ~ / 10 μl μl 25 ng /μl 1. 0 μl 10 PCR Buffer 2. 0 μ Mg mmol /L 1. 6 μl Taq 5 U /μl 0. 2 μl dntp 10 mmol /L 0. 2 μl ddh 2 O μl 20 μl s s 72 1 min min D μl MseⅠ 25 ng /μl 0. 8 μl EcoRⅠ 10 ng /μl 0. 8 μl 10 PCR Buffer1. 0 μ Mg mmol /L 0. 8 μl Taq 5 U /μl 0. 1 μl dntp 10 mmol /L 0. 2 μl ddh 2 O 5. 3 μl 10 μl 95 3 min s s 72 1 min s s 72 1 min min 14 PCR 8% CANON SX30 IS NTSYS UPGMA b Eb 1-Da 1-B 1-C traits by space-induced mutation 3 1-C ~ g ~ kg /hm 2 D15 EcoRⅠ + A 10 ng /μl 1. 0 μl MseⅠ + C 122 / kg / hm 2 D15 06ms198 05N01 D ms198 D15 A. 11 B C. D. a 11 b 11 E. a A. Yuzhi No. 11 capsule type of CK B C. Capsule type after space-induced mutation D. a was flower type after space-induced mutation of Yuzhi No. 11 b was flower type of Yuzhi No. 11 CK E. a was leaf type of Yuzhi No. 11 CK b was leaf type after space-induced mutation of Yuzhi No. 11. Fig. 1 The effect on sesame agronomic SP4 SP3 SP SP3 SP N ms D15 06ms198 D15

4 230 华北农学报 28 卷 06ms198 D15 2 SP4 SP3 SP ms198 1 Tab. 1 SP4 The main yield character in SP4 generation Varieties / kg /hm 2 Yield /cm Plant height /cm /cm / /g First capsule height Yellow apex length Number of capsules 1000-seed per plantweight 11 CK Yuzhi NO N01 CK Zhengzhi N D15 CK Zhengzhi D ms198 CK Zhengzhi ms CK Japan black sesame D % ~ % 06ms198

5 1 期高桐梅等 : 芝麻太空诱变效应及 AFLP 标记检测 231 Tab. 2 2 SP3 SP4 The comparison of yield traits between SP3 and SP4 generations of different sesame variety Mean SD CV SP3 SP3 SP4 SP4 Varieties Trait SP3 SP4 SP3 SP4 CK Mutation CK Mutation Yuzhi No N Zhengzhi N D Zhengzhi D ms Zhengzhi ms Japan black sesame Tab. 3 Varieties 3 The comparison of disease resistance traits of different sesame variety /% Survived rate CK Mutation /% Compared with CK 11 Yuzhi No N D ms ** Japan black sesame ** * ** Note * and ** mean significant difference at and level respectively ~ % ~ AFLP 3-A SP NTSYS

6 232 华北农学报 28 卷 ~ ~ % % ~ ~ B ~ DNA Fig. 2 PCR SDS-PAGE results of PCR products of parts 3 Fig. 3 3 AFLP Cluster analysis of AFLP for aerospace mutation 53 ~ - 36 cm 77 ~ - 30 cm 16 ~ - 19 cm 122 ~ / ~ g ~ kg /hm SP3 SP4 SP

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