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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (3): 335-343.doi: 10.3724/SP.J.1006.2019.82035

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Introducing qSS-9 Kas into Ningjing 4 by molecular marker-assisted selection to improve its seed storage ability

Ping ZHANG1,Yi-Mei JIANG1,Peng-Hui CAO1,Fu-Lin ZHANG1,Hong-Ming WU1,Meng-Ying CAI1,Shi-Jia LIU1,Yun-Lu TIAN1,Ling JIANG1,*(),Jian-Min WAN1,2   

  1. 1 State Key Laboratory of Crop Genetics and Germplasm Enhancement / Key Laboratory of Biology, Genetics and Breeding of Japonica Rice in Mid-lower Yangtze River, Ministry of Agriculture / Research Center of Jiangsu Plant Gene Engineering, Nanjing Agricultural University, Nanjing 210095, Jiangsu, China
    2 National Key Facility for Crop Gene Resources and Genetic Improvement / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2018-07-06 Accepted:2018-12-24 Online:2019-03-12 Published:2019-01-03
  • Contact: Ling JIANG E-mail:jiangling@njau.edu.cn
  • Supported by:
    This study was supported by the National Major Project for Developing New GM Crops(2016ZX08001006);the Key Science and Technology Project of Jiangsu Province(BE2018388);the Key Science and Technology Project of Jiangsu Province(BE2017368);Jiangsu Collaborative Innovation Center for Modern Crop Production

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Abstract:

In this paper, the chromosome segment substitution line SL36 was used as the donor parent with the Kasalath allele at the qSS-9 locus. Ningjing 4, a commercial cultivar with ideal agronomic traits, was used as a recurrent parent, through continuous self-pollination and backcrossing for four consecutive years. Four molecular markers Y-10, Y-11, Y-14, Y-13 closely linked to qSS-9 were used to screen genotypes, and molecular marker-assisted selection (MAS) was used for seed storage ability breeding for Ningjing 4. Through genetic improvement, we obtained inheritable lines with high seed storage ability. Most agronomic traits of the lines were nearly the same as those of Ningjing 4. These lines showed significantly higher germination rate, lower malondialdehyde content and more obvious TTC staining effects under artificial aging and natural aging conditions compared with Ningjing 4, indicating that the new lines with qSS-9 Kas have high seed storage ability.

Key words: rice, chromosome segment substitution line, molecular marker-assisted selection, seed storage ability, genetic improvement

Fig. 1

Graphical genotypes of chromosome segment substitution line SL36 (A) and backcross breeding scheme between Ningjing 4 and SL36 (B)"

Table 1

Markers used for qSS-9 mark-assisted selection in this study "

标记
Marker		 正向引物
Forward primer (5°-3°)		 反向引物
Reverse primer (5°-3°)
Y-10 ACCTCAAGTTTTCCTATTAGT TAGAGTGACCTGCTAATGAG
Y-11 ACGATTAGTTCAGTCCTTACAC AACGGCTCAACGATCAGTAC
Y-14 AAAAAGGATGGGAAACTGACC TTTGAACTCAGTACCTTGGGG
Y-13 AAAAAGGATGGGAAACTGACC TACAAATAATCCCGATGCCG

Fig. 2

Germination rates and tetrazolium assay of parents and F1 after natural aging for 18 months A : Germination rates of parents and F1; B,C: Seed germination and tetrazolium assay of parents SL36, Ningjing 4 and F1 (from left to right). ** means significant at the 0.01 probability level compared with the germination rate of Ningjing 4."

Fig. 3

Genotype identification of parents and BC5F2 by molecular markers Y-10, Y-11, Y-14, and Y-13 close linked to qSS-9 A: Genotype identification of parents by molecular markers Y11 and Y14 close linkage to qSS-9; 1: DNA marker; 2-5, 6-9, 10-13, and 14-17 indicate genotype of markers Y-10, Y-11, Y-14, and Y-13 respectively; 2, 6, 10, and 14 indicate genotype of Nipponbare; 3, 7, 11, and 15 indicate genotype of Ningjing 4; 4, 8, 12, and 16 indicate genotype of SL36; 5, 9, 13, and 17 indicate genotype of Kasalath, respectively. B: Genotype identification of Ningjing 4, SL36 and BC5F2 by molecular markers Y11 and Y14 close linked to qSS-9; 1: DNA marker; 2-6 : Genotype of Ningjing 4, SL36 and BC5F2 under marker Y-10; 7-11: Genotype of Ningjing 4, SL36 and BC5F2 under marker Y-11; 12-16: Genotype of Ningjing 4, SL36 and BC5F2 under marker Y-14; 17-21: Genotype of Ningjing 4, SL36 and BC5F2 under marker Y-13, respectively."

Fig. 4

Germination rate and MDA contents in seeds of Ningjing 4, SL36, and progenies of different generations A: Contents of MDA in seeds of Ningjing 4, SL36, and BC3F2 after natural aging for one year; B: Germination rate of Ningjing 4, SL36 and BC3F2 after natural aging forone year and two years; C: Germination rates of parents and BC5F1 after artificial aging for 35 days; D: Seed germination of BC5F2 and Ningjing 4 after accelerated aging for 30 d and storage at 40°C and 80% RH; E-H: Seed viability of the control after aging treatment; E, F, G, and H represent Ningjing 4, Y3059-1-7, Y3059-1-8, Y3059-1-9, respectively. Significant differences relative to the control were assessed by t-tests. * and **, significant at P = 0.05 and P = 0.01, respectively. Values are means ± SD (n = 2)."

Fig. 5

Comparison of Ningjing 4 and Y3059 in tetrazolium assays, apparent amylose content and plant types A: Tetrazolium assay of Ningjing 4 and Y3059; From left to right represent Ningjing 4, Y3059-1-7, Y3059-1-8, Y3059-1-9, respectively; B: The content of TTCH of Y3059 and Ningjing 4; C: The change of apparent amylose content in Ningjing 4 and Y3059; D: Plant type of Ningjing 4 and BC5F2 at heading; E: Plant type of Ningjing 4 and BC5F2 at 35 days after heading; Significant differences relative to the control were assessed by t-tests. * and **, significant at P = 0.05 and P = 0.01, respectively. Values are means ± SD (n = 2)."

Table 2

Comparison of agronomic traits between Ningjing 4 and Y3059"

抽穗期
HD (d)		 株高
PH (cm)		 分蘖数
NT		 结实率
SSR (%)		 千粒重
TGW (g)		 单株产量
YPP (g)
宁粳4号Ningjing 4 87 ± 3 95 ± 4 11 ± 2 80.74 ± 0.02 25.50 ± 0.20 23 ± 0.18
BC5F2 (Y3059) 85 ± 3 107.5 ± 2.2** 13 ± 3* 83.08 ± 0.06 25.65 ± 0.90 22 ± 0.20
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