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Acta Agron Sin ›› 2009, Vol. 35 ›› Issue (9): 1620-1627.doi: 10.3724/SP.J.1006.2009.01620

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

Cloning and Expression Analysis of cMDH Gene Related to Cytoplasmic Male Sterile Wheat  with Aegilops kotschyi Aytoplasm

ZHANG Long-Yu,LI Hong-Xia,ZHANG Gai-Sheng*,WANG Jun-Sheng,HAN Yan-Fen,YUAN Zheng-Jie,NIU Na,MA Shou-Cai   

  1. Key Laboratory of Crop Heterosis of Shaanxi Province,Northwest A&F University/Wheat Breeding Engineering Research Center,Ministry of Education,Yangling 712100,China
  • Received:2009-02-24 Revised:2009-04-08 Online:2009-09-12 Published:2009-07-03
  • Contact: ZHANG Gai-Sheng，E-mail: zhanggsh@public.xa.sn.cn

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Abstract:

Male sterility withAegilops kotschyi cytoplasmhas a great application potential in hybrid wheat (Triticum aestivum L.) breeding for its stable sterility and broad-spectrum of restoring gene resources. To furtherreveal the genetic mechanism of male sterile with Ae. kotschyi cytoplasm, we employed a male sterile line ms (Kots)-90-110(A) and its near isogenic line BC4F1 (fertility restored by rk5451) to construct sterile and fertile cDNA libraries by suppression subtractive hybridization (SSH) in binucleate stage of anther development. Comparative analysis of differentially expressed EST sequences revealed that one EST highly similar to cytosolic malate dehydrogenases gene was identified from the fertile SSH-cDNA library. Then, The EST sequence was used as a querying probe to blast the Genbank databases. Based on the assembled homologous cDNA sequence, both cDNA and DNA sequences encoding a cytosolic malate dehydrogenases were isolated and characterized by PCR and sequence analysis. Furthermore, expression characteristics of the gene between male sterile and fertile anthers were analyzed via real-time PCR. In this study, the cDNA sequence was 1213 bp in length and the open reading frame encoded a peptide of 333 amino acids. The DNA sequence was 2908 bp in length, which contained seven extrons and six introns. According to expression analysis, the expression of this gene in fertile anthers was much higher than that in sterile anthers at binucleate and trinucleate stage during anther development. The trend of MDH activity was consistent with the quantitative results between fertility and sterility. Therefore, the gene is conjectured to be an early expression gene and its down-regulated expression may affect energy supply during stamen growth in sterile line anthers resulting in male sterility in wheat.

Key words: Wheat, Cytosolic malate dehydrogenases gene, Cloning, Real-time PCR

[1]Ocheretina O, Scheibe R. Cloning and sequence analysis of cDNAs encoding plant cytosolic MDH. Gene, 1997, 199: 145-148

[2]Bryan J K. Biosynthesis and regulation of anino acid. In: Boner J, Varner J E, eds. Plant Biochemistry, 3nd ed. New York: Academic Press, 1976. pp 351-371

[3]Mauseth J D. Botany: An Introduction to Plant Biology. Florida: Saunders College Publishing, 1991. pp 272-296

[4]Hu J-G(胡建广), Zhao X-S(赵相山), Liu J(刘军), Yuan Z-Q(袁自强), Yang J-S(杨金水). Isolation and characterization of a cDNA encoding maize cytosolic malate dehydrogenase. Acta Bot Sin (植物学报), 1999, 41(1): 40-44 (in Chinese with English abstract)

[5]Jiang R-H(姜瑞华), Lin C-F(林长发), Shen G-A(申国安), Qian M(钱旻), Wang D(王东), Du X-L(杜喜玲), Huang J(黄骥), Yang J-S(杨金水). Cloning of rice malate dehydrogenase and expression in E. coli. J Fudan Univ (Nat Sci)(复旦学报自然科学版), 2002, 41(1): 67-69 (in Chinese)

[6]Lin Z B, Xu Y, Ma Q H, D Y. Cloning and evolutionary analysis of a partial cytosolic malate dehydrogenase cDNA from wheat. J Agric Biotechnol, 2004, 12: 38-42

[7]Zhang G-S(张改生), Yang T-Z(杨天章). A preliminary study on the male sterile lines of wheat with Ae. ventricosa, Ae. kotscgyi and Ae. variabilis cytoplasms. Acta Agron Sin (作物学报), 1989, 15(1): 1-10 (in Chinese with English abstract)

[8]Li H-X(李红霞), Zhang L-Y(张龙雨), Zhang G-S(张改生), Niu N(牛娜), Zhu Z-W(朱展望). Construction on SSH library from fertility-related genes of male sterility of wheat with Aegilops kotschyi cytoplasm. Acta Agron Sin (作物学报), 2008, 34 (6): 965-971 (in Chinese with English abstract)

[9]Zhang C-Y(张驰宇), Xu S-G(徐顺高), Huang X-X(黄新祥). A novel and convenient relative quantitative method of fluoresce-nce real time RT-PCR assay based on slope of standard curve. Prog Biochem Biophys (生物化学与生物物理进展), 2005, 32 (9): 883-887 (in Chinese with English abstract)

[10] Ou-Yang G-C(欧阳光察). Determination of malate dehydrogenase activity. In: Shanghai Society for Plant Physiology. Laboratory Manual for Plant Physiology (植物生理学实验手册). Shanghai: Shanghai Scientific and Technical Publishers, 1985. pp 179-181 (in Chinese)

[11]Zhu G-L(朱广廉), Zhong H-W(钟诲文), Zhang A-Q(张爱琴). Experimental Guide for Plant Physiological (植物生理学实验). Beijing: Peking University Press, 1990. pp 32-34 (in Chinese)

[12] Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method. Methods, 2001, 25: 402-408

[13] Liu W, Saint D A. A new quantitative method of real time reverse transcription polymerase chain reaction assay based on simulation of polymerase chain reaction kinetics. Anal Biochem, 2002, 302: 52-59

[14] Giulietti A, Overbergh L, Valckx D, Decallonne B, Bouillon R, Mathieu C. An overview of real-time quantitative PCR: Applications to quantify cytokine gene expression. Methods, 2001, 25: 386-401

[15] Heber U. Metabolite exchange between chloroplasts and cytoplasm. Annu Rev Plant Physiol, 1974, 25: 393-421

[16] Mascarenhas J P. Gene activity during pollen development. Aunu Rev Plant Physiol Plant Mol Biol, 1990, 41: 317-338

[17] Gong H-W(龚宏伟), Ma L-J(马翎健), He B-R(何蓓如), Fan C-Y(范春燕), Wu H(武涵), Li W(李薇). Changes of RNase and protein content in the fertility sensitive period of wheat male sterile lines. Triticeae Crops (麦类作物学报), 2008, 28(1): 31-34 (in Chinese with English abstract)

[18] Wu F(吴峰), Liu Y-M(刘玉梅), Sun D-L(孙德岭), Wu F(吴锋), Chang C-T(常彩涛). Biochemical traits of cytoplasmic male sterile and maintainer lines in hot pepper. Tianjin Agric Sci (天津农业科学), 2008, 14(2): 50-52 (in Chinese with English abstract)

[19] Li P(李平), Liu H-X(刘鸿先), Wang Y-R(王以柔), Zeng S-X(曾韶西), Li M-R(李美如), Zhang X(张旭), Lin D-X(林道宣), Chen Z-M(陈钊明), Liu Y-Z(刘彦卓), Kong Q-N(孔清霓). The fertility expression of dual-purpose genic male sterile line indica rice Pei’ai 64S: Changes of NAD+-MDH and AP isoenzyme during spikelet development. Chin J Rice Sci (中国水稻科学), 1997, 11(2): 83-88 (in Chinese with English abstract)

[20] Zhang M-Y(张明永), Liang C-Y(梁承邺), Huang Y-W(黄毓文), Liu H-X(刘鸿先). Comparison of respiratory pathways of CMS and its maintainer rice (Oryza stativa L.). Acta Phytophysiol Sin (植物生理学报), 1998, 24 (1): 55-58 (in Chinese with English abstract)

[21] Liang C-Y(梁承邺), Chen X-F(陈贤丰), Sun G-C(孙谷畴), Huang Y-W(黄毓文), Lin Z-F(林植芳). Some biochemical metabolic characters in Nongken 58s anthers of Hubei photoperiod sensitive genic male-sterile rice. Acta Agron Sin (作物学报), 1995, 21(1): 64-70 (in Chinese with English abstract)
[22] Yao Y-Q(姚雅琴), Zhang G-S(张改生), Liu H-W(刘宏伟), Wang J-W(王军卫), Liu H-M(刘红梅). Cytomorphology and cytochemical localization of K-type and T-type cytoplasmic male sterile pollens in wheat. Sci Agric Sin (中国农业科学), 2002, 35(2): 123-126 (in Chinese with English abstract)
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