Welcome to Acta Agronomica Sinica，

Acta Agron Sin ›› 2010, Vol. 36 ›› Issue (10): 1634-1641.doi: 10.3724/SP.J.1006.2010.01634

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

Cloning and SNP Analysis of TT1 Gene in Brassica juncea

YAN Ming-Li1,2,LIU Xian-Jun2,GUAN Chun-Yun2,LIU Li-Li1,LU Ying1,2,LIU Zhong-Song2,*   

  1. 1 School of Biology, Hunan University of Science and Technology, Xiangtan 411201, China; 2 Oilseed Research Institute, Hunan Agricultural University, Changsha 410128, China
  • Received:2010-04-05 Revised:2010-05-03 Online:2010-10-12 Published:2010-08-04
  • Contact: LIU Zhong-song,E-mail:zsliu48@sohu.com;Tel:0731-84617381

PDF

1893

Cited

7

Abstract: The quality of rapeseed can be further improved through the development of yellow-seeded cultivars, which are known to contain less lignin and more proteins, both enhancing the dietary feed value. The TT1 gene encodes a WIP domain protein with Zn-finger, which is essential for seed coat development and organ color in Arabidopsis, its mutation causes transparent testa. In order to study molecular mechanism of seed coat color in rapeseed, TT1 gene was cloned from Brassica juncea using homology-based cloning and RACE (rapid-amplification of cDNA ends) strategy. A modified allele-specific PCR procedure developed for assaying single nucleotide polymorphisms of TT1 gene in Brassica juncea. In this study, cloning and characterization of the orthologous TT1 gene from Brassica juncea were reported. The full length DNA sequence of TT1 gene was 2 197 bp with one intron. The TT1 cDNA sequence was 1 412 bp in length, 903 bp of which was open reading frames (ORF) encoding a deduced polypeptide of 300 amino acids with a predicted molecular weight of 33.97 kD and an isoelectric point of 6.99. The above-mentioned genomic sequences showed 99% identity with the TT1 gene from Brassica napus, and 85% identity with the TT1 gene from Arabidopsis. RT-PCR analysis showed that TT1 expressed in the seed coats of both the black- and the yellow-seeded B. juncea lines. Comparison of sequences of Sichuan Yellow (yellow seed) and purple-leaf mustard (black seed) revealed nucleotide variation at the eight sites which all are located within the coding region of the gene. However, the mutations of the TT1 gene at these sites had no effect on the seed coat color in B. juncea through comparing the sequences of NILB and purple-leaf mustard. The allele-specific PCR of the TT1 gene could distinguish purple-leaf mustard from Sichuan Yellow.

Key words: Brassica juncea, TT1, Cloning, Single nucleotide polymorphism

[1] Rashid A, Rakow G, Downey R K. Development of yellow seeded Brassica napus through interspecific crosses. Plant Breed, 1994, 112: 127–134
[2] Rahman M H, Joersho M , Poulsen M H. Development of yellow-seeded Brassica napus of double low quality. Plant Breed, 2001, 120: 473–478
[3] Vera C L, Woods D L. Isolation of independent gene pairs at two loci for seed coat color in Brassica juncea. Can J Plant Sci, 1982, 62: 47–50
[4] Yan M L, Liu Z S, Guan C Y, Chen S Y, Yuan M Z, Liu X J. Inheritance and molecular markers for the seed coat color in Brassica juncea. Front Agric China, 2009, 3: 1–6
[5] Lepiniec L, Debeaujon I, Routaboul J M, Baudry A, Pourcel L, Nesi N, Caboche M. Genetics and biochemistry of seed flavonoids. Ann Rev Plant Biol, 2006, 57: 405–430
[6] von Wettstein D. From analysis of mutants to genetic engineering. Ann Rev Plant Biol, 2007, 58: 1–19
[7] Chai Y R, Lei B, Huang H L, Li J N, Yin J M, Tang Z L, Wang R, Chen L. TRANSPARENT TESTA 12 genes from Brassica napus and parental species: Cloning, evolution, and differential involvement in yellow seed trait. Mol Genet Genom, 2009, 281: 109–123
[8] Debeaujon I, Leon-Kloosterziel K M, Koornneef M. Influence of the testa on seed dormancy, germination and longevity in Arabidopsis. Plant Physiol, 2000, 122: 403–413
[9] Sharma S B, Dixon R A. Metabolic engineering of proanthocyanidins by ectopic expression of transcription factors in Arabidopsis thaliana. Plant J, 2005, 44: 62–75
[10] Sagasser M, Lu G H, Hahlbrock K, Weisshaar B. A. thaliana TRANSPARENT TESTA 1 is involved in seed coat development and defines the WIP subfamily of plant zinc finger proteins. Genes Dev, 2002, 16: 138–149
[11] Nesi N, Lucas M O, Auger B, Baron C, Lecureuil A, Guerche P, Kronenberger J, Lepiniec L, Debeaujon I, Renard M. The promoter of the Arabidopsis thaliana BAN gene is active in proanthocyanidins-accumulating cells of Brassica napus seed coat. Plant Cell Rep, 2009, 28: 601–617
[12] Liu X-J(刘显军), Yuan M-Z(袁谋志), Guan C-Y(官春云), Chen S-Y(陈社员), Liu S -Y(刘淑艳), Liu Z-S(刘忠松). Inheritance, mapping, and origin of yellow-seeded trait in Brassica juncea. Acta Agron Sin (作物学报), 2009, 35(5): 839−847 (in Chinese with English abstract)
[13] Yan M-L(严明理), Liu Z-S(刘忠松), Guan C-Y(官春云), Chen S-Y(陈社员), Liu X-J(刘显军). Method for high-quality RNA isolation from the seeds and their testa of Brassica species. Biotech Bull生物技术通报), 2007, (6): 97–100 (in Chinese with English abstract) (
[14] Dellaporta S L, Wood J, Hicks J B. A plant DNA minipreparation: version II. Plant Mol Biol Rept, 1983, 1: 19–21
[15] Yan M-L(严明理), Liu X-J(刘显军), Liu Z-S(刘忠松), Guan C-Y(官春云), Yuan M-Z(袁谋志), Xiong X-H(熊兴华). Cloning and expression analysis of the dihydroflavonol 4-reductase gene in Brassica juncea. Acta Agron Sin (作物学报), 2008, 34(1): 1–7 (in Chinese with English abstract)
[16] Drenkard E, Richter B G, Rozen S, Stutius L M, Angell N A, Mindrinos M, Cho R J, Oefner P J, Davis R W, Ausubel F M. A simple procedure for the analysis of single nucleotide polymorphisms facilitates map-based cloning in Arabidopsis. Plant Physiol, 2000, 124: 1483–1492
[17] Hayashi K, Hashimoto N, Daigen M, Ashikawa I. Development of PCR-based SNP markers for rice blast resistance genes at the Pi-z locus. Theor Appl Genet, 2004, 108: 1212–1220
[18] Lu X-C(路小春). Molecular Cloning of Transparent Testa (BnTT1) Gene Family Encoding WIP-Zinc Finger Proteins and Differential Expression between Yellow- and Black-seeded Lines of Brassica napus. MS Dissertation of Yunnan Agricultural University, 2006 (in Chinese with English abstract)
[19] Wolfe S A, Nekludova L, Pabo C O. DNA recognition by Cys2His2 zinc finger proteins. Ann Rev Biophys Biomol Struct, 2000, 29: 183–212
[20] Mackay J P, Crossley M. Zinc fingers are sticking together. Trends Biochem Sci, 1998, 23: 1–4
[21] Laity J H, Lee B M, Wright P E. Zinc finger proteins: New insights into structural and functional diversity. Curr Opin Struct Biol, 2001, 11: 39–46
[22] Auger B, Baron C, Lucas M O, Vautrin S, Berges H, Chalhoub B, Fautrel A, Renard M, Nesi N. Brassica orthologs from BANYULS belong to a small multigene family, which is involved in procyanidin accumulation in the seed. Planta, 2009, 230: 1167–1183
[23] Wu Y H, Xiao L, Wu G , Lu C M. Cloning of fatty acid elongase 1 gene and molecular identification of A and C genome in Brassica species. Sci China (Ser C-Life Sci), 2007, 50: 343–349
[24] Winkel-Shirley B. Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol, 2001, 126: 485–493
[25] Lysak M A, Berr A, Pecinka A, Schmidt R, McBreen K, Schubert I. Mechanisms of chromosome number reduction in Arabidopsis thaliana and related Brassicaceae species. Proc Nat Acad Sci USA, 2006, 103: 5224–5229
[26] Lysak M A, Cheung K, Kitschke M, Bures P. Ancestral chromosomal blocks are triplicated in Brassiceae species with varying chromosome number and genome size. Plant Physiol, 2007, 145: 402–410
[1] CUI Lian-Hua, ZHAN Wei-Min, YANG Lu-Hao, WANG Shao-Ci, MA Wen-Qi, JIANG Liang-Liang, ZHANG Yan-Pei, YANG Jian-Ping, YANG Qing-Hua. Molecular cloning of two maize (Zea mays) ZmCOP1 genes and their transcription abundances in response to different light treatments [J]. Acta Agronomica Sinica, 2022, 48(6): 1312-1324.
[2] ZHOU Hui-Wen, QIU Li-Hang, HUANG Xing, LI Qiang, CHEN Rong-Fa, FAN Ye-Geng, LUO Han-Min, YAN Hai-Feng, WENG Meng-Ling, ZHOU Zhong-Feng, WU Jian-Ming. Cloning and functional analysis of ScGA20ox1 gibberellin oxidase gene in sugarcane [J]. Acta Agronomica Sinica, 2022, 48(4): 1017-1026.
[3] XIE Qin-Qin, ZUO Tong-Hong, HU Deng-Ke, LIU Qian-Ying, ZHANG Yi-Zhong, ZHANG He-Cui, ZENG Wen-Yi, YUAN Chong-Mo, ZHU Li-Quan. Molecular cloning and expression analysis of BoPUB9 in self-incompatibility Brassica oleracea [J]. Acta Agronomica Sinica, 2022, 48(1): 108-120.
[4] TANG Rui-Min, JIA Xiao-Yun, ZHU Wen-Jiao, YIN Jing-Ming, YANG Qing. Cloning of potato heat shock transcription factor StHsfA3 gene and its functional analysis in heat tolerance [J]. Acta Agronomica Sinica, 2021, 47(4): 672-683.
[5] YUE Jie-Ru, BAI Jian-Fang, ZHANG Feng-Ting, GUO Li-Ping, YUAN Shao-Hua, LI Yan-Mei, ZHANG Sheng-Quan, ZHAO Chang-Ping, ZHANG Li-Ping. Cloning and potential function analysis of ascorbic peroxidase gene of hybrid wheat in seed aging [J]. Acta Agronomica Sinica, 2021, 47(3): 405-415.
[6] LI Jing-Lin, LI Jia-Lin, LI Xin-Peng, AN Bao-Guang, ZENG Xiang, WU Yong-Zhong, HUANG Pei-Jing, LONG Tuan. Creation and combining ability analysis of recessive genic sterile lines with a new ptc1 locus in rice [J]. Acta Agronomica Sinica, 2021, 47(11): 2173-2183.
[7] YANG Qin-Li, YANG Duo-Feng, DING Lin-Yun, ZHANG Ting, ZHANG Jun, MEI Huan, HUANG Chu-Jun, GAO Yang, YE Li, GAO Meng-Tao, YAN Sun-Yi, ZHANG Tian-Zhen, HU Yan. Identification of a cotton flower organ mutant 182-9 and cloning of candidate genes [J]. Acta Agronomica Sinica, 2021, 47(10): 1854-1862.
[8] HE Xiao, LIU Xing, XIN Zheng-Qi, XIE Hai-Yan, XIN Yu-Feng, WU Neng-Biao. Molecular cloning, expression, and enzyme kinetic analysis of a phenylalanine ammonia-lyase gene in Pinellia ternate [J]. Acta Agronomica Sinica, 2021, 47(10): 1941-1952.
[9] PENG Bo,ZHAO Xiao-Lei,WANG Yi,YUAN Wen-Ya,LI Chun-Hui,LI Yong-Xiang,ZHANG Deng-Feng,SHI Yun-Su,SONG Yan-Chun,WANG Tian-Yu,LI Yu. Genome-wide association studies of leaf orientation value in maize [J]. Acta Agronomica Sinica, 2020, 46(6): 819-831.
[10] WANG Rui-Li,WANG Liu-Yan,YE Sang,Gao Huan-Huan,LEI Wei,WU Jia-Yi,YUAN Fang,MENG Li-Jiao,TANG Zhang-Lin,LI Jia-Na,ZHOU Qing-Yuan,CUI Cui. QTL mapping of seed germination-related traits in Brassica napus L. under aluminum toxicity stress [J]. Acta Agronomica Sinica, 2020, 46(6): 832-843.
[11] Tong-Hong ZUO, He-Cui ZHANG, Qian-Ying LIU, Xiao-Ping LIAN, Qin-Qin XIE, Deng-Ke HU, Yi-Zhong ZHANG, Yu-Kui WANG, Xiao-Jing BAI, Li-Quan ZHU. Molecular cloning and expression analysis of BoGSTL21 in self-incompatibility Brasscia oleracea [J]. Acta Agronomica Sinica, 2020, 46(12): 1850-1861.
[12] ZHANG Hong-Juan,LI Yu-Ying,MIAO Li-Li,WANG Jing-Yi,LI Chao-Nan,YANG De-Long,MAO Xin-Guo,JING Rui-Lian. Transcription factor gene TaNAC67 involved in regulation spike length and spikelet number per spike in common wheat [J]. Acta Agronomica Sinica, 2019, 45(11): 1615-1627.
[13] XUE Xiao-Meng,LI Jian-Guo,BAI Dong-Mei,YAN Li-Ying,WAN Li-Yun,KANG Yan-Ping,HUAI Dong-Xin,LEI Yong,LIAO Bo-Shou. Expression profiles of FAD2 genes and their responses to cold stress in peanut [J]. Acta Agronomica Sinica, 2019, 45(10): 1586-1594.
[14] Huan TAN,Yu-Hui LIU,Li-Xia LI,Li WANG,Yuan-Ming LI,Jun-Lian ZHANG. Cloning and Functional Analysis of R2R3 MYB Genes Involved in Anthocyanin Biosynthesis in Potato Tuber [J]. Acta Agronomica Sinica, 2018, 44(7): 1021-1031.
[15] Jing DONG,Xiao-Ping LU,Kun-Ming ZHANG,Chun-Lei XUE,Rui-Xia ZHANG. Analysis of SNP and Allele-specific Expression in Transcriptome of Sorghum bicolor × Sorghum sudanense and Their Parents [J]. Acta Agronomica Sinica, 2018, 44(12): 1809-1817.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd