Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (23): 4759-4769.doi: 10.3864/j.issn.0578-1752.2012.23.002

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

Cloning and Expression Analysis of A Putative B Class MADS-box Gene of AcPI in Onion

 LI  Hong-You, WANG  Chan, LI  Li-Lin, WANG  Yong-Qin, ZHAO  Rui   

  1. 1.College of Horticulture, Shenyang Agricultural University, Shenyang 110161;
    2.Beijing Vegetable Research Center, Beijing Academy of Agriculture and Forestry Sciences/Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (North China), Ministry of Agriculture, Beijing 100097;
    3.School of Life Science, Shanxi University, Taiyuan 030005
  • Received:2012-08-15 Online:2012-12-01 Published:2012-09-14

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Abstract: 【Objective】The aim of this study is to clone the PI/GLO subfamily MADS-box gene of class B of the floral identity in onion, to analyze its sequence characteristics and the temporal and spatial expression pattern. The results will provide a foundation for further dissection of the molecular and genetics mechanisms of floral organ development in onion in detail.【Method】Using the total RNA from the floral buds of onion (Allium cepa) as the template, we design the degenerate primers based on homology cloning strategy, and then the full-length cDNA sequence of AcPI was obtained through a combined reverse transcription-PCR (RT-PCR) and RACE techniques strategy. The bioinformatics methods were used to analyze the sequence characteristics of this gene and the combination of RT-PCR and Real-time PCR methods were used to investigate the temporal and spatial expression pattern of AcPI gene.【Result】The cloned full-length cDNA of AcPI was 931 bp in length, containing a 615 bp open reading frame (ORF) which encodes 205 amino acids. Protein alignment analysis indicated that AcPI protein had typical structure domains of plants MADS-box protein, containing a MADS domain and a K domain. Protein sequence identity search showed that AcPI is similar to NTPI (Narcissus tazetta var. chinensis), HoMADS2 (Hyacinthus orientalis) and GLO (Antirrhinum majus) with similarities of 78%, 75% and 52% respectively. Phylogenetic tree reconstructed clearly showed that AcPI protein was classed into PI subgroup of B class MADS-box protein family. RT-PCR analysis indicated that expression of the AcPI gene was restricted to the flowers as its mRNA was not detected in vegetative tissues such as roots, stems, leaves. Real-time PCR analysis showed that AcPI gene mainly expressed in the first, second and third whorls of flower organ and only weak expression was detected in the fourth whorls of flower organ in all three flower bud developmental stages. Furthermore, the expression level of AcPI was increasing in all flower organs except in the first whorl flower organ is first increases and then decreases in the throughout growing process of flower buds.【Conclusion】The extended expression of AcPI in the first whorl floral organ supported the modified ABC model by van Tunen, but it also expressed in the fourth whorl floral organ, which showed that AcPI gene may play an important role in the development of carpels.

Key words: onion , floral organs , MADS-box gene , gene cloning , expression analysis

[1]Bowman J L, Smyth D R, Meyerowitz E M. Genes directing flower development in Arabidopsis. The Plant Cell, 1989, 1: 37-52.

[2]Meyerowitz E M, Bowman J L, Brockman L L, Drews G N, Jack T, Sieburth L E, Weigel D. A genetic and molecular model for flower development in Arabidopsis thaliana. Development, 1991, 1: 157-167.

[3]Meyerowitz E M, Symth D R, Bowman J L. Abnormal flowers and pattern formation in floral development. Development, 1989, 106: 209-217.

[4]Schwarz-Sommer Z, Huijser P, Nacken W, Saedler H, Sommer H. Genetic control of flower development by homeotic genes in Antirrhinum majus. Science, 1990, 250: 931-936.

[5]Coen E S, Meyerowitz E M. The war of the whorls: genetic interactions controlling flower development. Nature, 1991, 353: 31-37.

[6]Angenentg G C, Franken J，Busscher M, van Dijken J, van Went J L, Dons H J, van Tunen A J. A novel class of MADS-box genes is involved in ovule development in petunia. The Plant Cell, 1995, 7(10): 1569-1582.

[7]Pelaz S, Ditta G S, Baumann E, Wisman E, Yanofsky M F. B and C floral organ identity functions require SEPALLATA MADS-box genes. Nature, 2000, 405(6783): 200-203.

[8]Ditta G, Pinyopich A, Robles P, Pelaz s, Yanofsky M F. The SEP4 gene of Arabidopsis thaliana functions in floral organ and meristem identity. Current Biology, 2004, 14(21): 1935-1940.

[9]van Tunen A J, Eikelboom W, Angenent G C. Floral organogenesis in Tulipa. Flowering Newsletter, 1993, 16: 33-38.  

[10]Kanno A, Saeki H, Kameya T, Saedler H, Theissen G. Heterotopic expression of class B floral homeotic genes supports a modified ABC model for tulip (Tulip gesneriana). Plant Molecular Biology, 2003, 52: 831-841.

[11]Winter K U, Weiser C, Kaufmann K, Bohne A, Kirchner C, Kanno A, Saedler H, Theissen G. Evolution of class B floral homeotic proteins: Obligate heterodi-merization originated from homodimerization. Molecular Biology and Evolution, 2002, 19: 587-596.

[12]Tzeng T Y, Yang C H. A MADS box gene from lily (Lilium longiflorum) is sufficient to generate dominant negative mutation by interacting with PI  (PISTILLATA) in Arabidopsis thaliana. Plant and Cell Physiology, 2002, 42: 1156-1168.

[13]Nakamura T, Fukuda T, Nakano M, Hasebe M, Kameya T, Kanno A. The modified ABC model explains the development of the petaloid perianth of Agapanthus praecox ssp. orientalis (Agapanthaceae) flowers. Plant Molecular Biology, 2005, 58: 435-445.

[14]Nakada M, Komatsu M, Ochiai T, Ohtsu K, Nakazono M, Nishizawa N K, Nitta K, Nishiyama R, Kameya T, Kanno A. Isolation of MaDEF from Muscari armeniacum and analysis of its expression using laser mcrodissection. Plant Science, 2006, 170: 143-150.

[15]Tsaftaris A S, Polidoros A N, Pasentsis K, Kalivas A. Tepal formation and expression pattern of B-class paleoAP3-like MADS-box genes in crocus (Crocus sativus L). Plant Science, 2006, 170: 238-246.

[16]Tsai W C, Kuoh C S, Chuang M H, Chen W H, Chen H H. Four DEF-like MADS box genes displayed distinct floral morphogenetic roles in Phalaenopsis orchid. Plant and Cell Physiology, 2004, 45: 831-844.

[17]Tsai W C, Lee P F, Chen H I, Hsiao Y Y, Wei W J, Pan Z J, Chuang M H, Kuoh C S, Chen W H, Chen H H. PeMADS6, a GLOBOSA/ PISTILLATA-like gene in Phalaenopsis equestris involved in petaloid formation, and correlated with flower longevity and ovary development. Plant and Cell Physiology, 2005, 46: 1125-1139.

[18]Xu Y, Teo L L, Zhou J, Kumar P P, Yu H. Floral organ identity genes in orchid Dendrobium crumenatum. The Plant Journal, 2006, 46: 54-68.

[19]Hsu H F, Yang C H. An orchid (Oncidium Gower Ramsey) AP3-like MADS gene regulates floral formation and initiation. Plant and Cell Physiology, 2002, 43: 1198-1209.

[20]Park J H, Ishikawa Y, Yoshida R, Kanno A, Kameya T. Expression of AODEF, a B-functional MADS-box gene, in stamens and inner tepals of dioecious species Asparagus officinalis L. Plant Molecular Biology, 2003, 51: 867-875.

[21]Park J H, Ishikawa Y, Ochiai T, Kanno A, Kameya T. Two GLOBOSA-like genes are expressed in second and third whorls of homochlamydeous flowers in Asparagus officinalis L. Plant Cell Physiology, 2004, 45: 325-332.

[22]夏  瑞, 陆旺金, 李建国. 一种改良的扩增cDNA 5′末端的方法. 园艺学报, 2008, 35(10): 1533-1538.

Xia R, Lu W J, Li J G. A modified method of amplifying the 5′-end cDNA sequence. Acta Horticulturae Sinica, 2008, 35(10): 1533-1538. (in Chinese)

[23]Kramer E M, Dorit R L, Irish V F. Molecular evolution of genes controlling petal and stamen development: duplication and divergence within the APETALA3 and PASTILLATA MADS-box gene lineages. Genetics, 1998, 149: 765-783.

[24]Krizek B A, Fletcher J C. Molecular mechanisms of flower development: an armchair guide. Nature Reviews Genetics, 2005, 6: 688-698.

[25]Münster T, Wingen L U, Faigl W, Werth S, Saedler H, Theissen G. Characterization of three GLOGOSA-like MADS-box genes from maize: evide-nce for ancient paralogy in one class of floral homeotic B-function genes of grasses. Gene, 2001, 262: 1-13.

[26]Yong-Yoon C, Seong-Ryong K, Hong-Gyu K, Yoo-Sun N, Park M C, Finkel D, Gynheung A. Characterization of two rice MADS box genes homologus to GLOBOSA. Plant Science, 1995, 109: 45-46.

[27]Becker A, Kaufmann K, Freialdenhoven A, Vincent C, Li M A, Saedler H, Theissen G. A novel MADS-box gene subfamily with a sister-group relationship to class B homeotic genes. Molecular Genetics and Genomics, 2002, 266: 942-950.

[28]陈小强, 王春国, 李秀兰, 宋文芹, 陈瑞阳. 大花蕙兰MADS-box基因ChMADS1的克隆及表达分析. 南开大学学报: 自然科学版, 2008, 41(6): 1-7.

Chen X Q, Wang C G, Li X L, Song W Q, Chen R Y. Cloning and sequence analysis of a novel MADS-box gene expression in the ovary from Cymbidium hybridium. Acta Scientiarum Naturalium Universitatis Nankaiensis, 2008, 41(6): 1-7.(in Chinese)

[29]Kanno A, Nakada M, Akita Y, Hirai M. Class B gene expression and the modified ABC model in nongrass monocots. The Scientific World Journal, 2007, 7: 268-279.

[30]Jover-Gil S, Candela H, Ponce M R. Plant microRNAs and development. International Journal of Developmental Biology, 2005, 49: 733-744.

[31]Theissen G, Saedler H. Floral quartets. Nature, 2001, 409: 469-471.
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