Isolation and Bioinformatics Analysis of Apomicxis SERK1 Genes in Malus

ZHANG Li-jie; MOHAMED Hamad; DONG Wen-xuan; GUO Su-man; MENG Qing-jiao

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Isolation and Bioinformatics Analysis of Apomicxis SERK1 Genes in Malus

1.
College of Forestry, Shenyang Agricultural University, Shenyang 110866, Liaoning, China
2.
College of Horticulture, Shenyang Agricultural University, Shenyang 110866, Liaoning, China
3.
Forestry Bureau of Huanren County, Benxi 117201, Liaoning, China

Received Date: 2015-06-26

Abstract

The plants Malus hupehensis var. pingyiensis Jiang (Pingyi Tiancha) and a hybrid strain 33# were employed as the experimental materials. By PCR techniques and primers which were designed from the CDS sequences of apple genome, the full-length cDNA sequences of SERK homologous genes were cloned, which were named as MhSERK1 and MhdSERK1 (GenBank accession No. JQ231273 and JQ231272), and then SERKs expressions were detected in different tissues and organs of Pingyi Tiancha and the hybrid strain through Real-time quantitative PCR method. The results showed that the length of coding region sequences about MhSERK1 and MhdSERK1 were 1 899 bp and 1 881 bp, which respectively encoded 632 and 626 amino acids. Compared with other plants, the amino acid sequence homology of SERK1 was 80% or more, especially with Longan grape (Vitaceae), which could reach 92.56%, and it also had a high homology with the model plant Arabidopsis thaliana and tobacco. The results of real-time quantitative PCR showed that the expression levels of SERK1 gene were different among the different tissues and organs of Pingyi Tiancha and hybrid strain, which was high expression levels in the ovary, very low expression levels in vegetative tissues, and the highest expression levels in the ovary of the flower bud of Pingyi Tiancha, indicating that SERK1 gene played an important role in the reproductive and developmental processes of Pingyi Tiancha and hybrid strain.
- Malus,
- SERK,
- homologue gene,
- bioinformatics analysis

References

[1]	束怀瑞.苹果学[M].北京:中国农业出版社,1999,210-235.
[2]	李光杰. 平邑甜茶SnRK1基因的克隆、表达及其功能的研究[D]. 泰安:山东农业大学, 2009.
[3]	史兴征. 平邑甜茶GLB1基因的克隆、表达及其功能的研究[D].泰安:山东农业大学, 2010.
[4]	张丽杰,苹果属无融合生殖相关SERK基因克隆与表达分析及遗传转化的研究[D]. 沈阳:沈阳农业大学, 2013.
[5]	Schmidt H. Contribution on the Breeding of Apomictic Apple Stocks 4. On the Inheritance of Apomixis[J]. Z Pflanzenzuchtg. 1977,78:312.
[6]	林盛华, 刘捍中, 蒲富慎. 苹果属植物不同倍性的杂种后代染色体数目观察[J]. 中国果树, 1989,(4): 15-16.
[7]	王颖, 商月惠, 王玉霞, 等.平邑甜茶与扎矮山定子杂交后代的胚胎发育特征研究[J]. 园艺学报, 2008,35(8):1093-1100.
[8]	王颖. 平邑甜茶(Malus hupehensis var.pingyiensis)的杂交后代生殖特性研究和花期子房抑制性消减文库构建[D]. 沈阳:沈阳农业大学, 2009.
[9]	林庆光, 崔百明, 彭明. SERK 基因家族的研究进展[J]. 遗传, 2007, 29(6): 681-687.
[10]	Schmidt E D, Guzzo F, Toonen M A, et al. A leucine-rich repeat containing receptor-like kinase marks somatic plant cells competent to form embryos[J]. Development, 1997, 124(10): 2049-2062.
[11]	Albrecht C, Russinova E, Hecht V, et al. The Arabidopsis thaliana SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASES1 and 2 control male sporogenesis[J]. Plant Cell, 2005,17(12): 3337-3349.
[12]	Colcombet J, Boisson-Dernier A, Ros-Palau R, et al. Arabidopsis SOMATIC EMBRYOGENESIS RECEPTOR KINASES1 and 2 are essential for tapetum development and microspore maturation[J]. Plant Cell, 2005, 17(12): 3350-3361.
[13]	Hecht V, Vielle-Calzada J P, Hartog M V, et al. The Arabidopsis somatic embryogenesis receptor kinase 1 gene is expressed in developing ovules and embryos and enhances embryogenic competence in culture[J]. Plant Physiol, 2001, 127(3): 803-816.
[14]	石雅丽, 张锐, 林芹, 等. 植物体细胞胚胎发生受体类蛋白激酶的生物学功能[J]. 遗传, 2012,34(5): 551-559.
[15]	邹冬梅,刘月学,张志宏,等. 草莓AP1同源基因的克隆与表达分析[J]. 中国农业科学,2012, 45(10):1972-1981.
[16]	Jun Ma, Yehua He, Chenghou Wu, et al. Cloning and molecular characterization of a SERK gene transcriptionally induced during somatic embryogenesis in Ananas comosus cv. Shenwan[J]. Plant Mol Biol Rep,2012, 30: 195-203.
[17]	Santos M D O, Romano E, Yotoko K S C, et al. Characterisation of the cacao somatic embryogenesis receptor-like kinase (SERK) gene expressed during somatic embryogenesis[J]. Plant Sci, 2005, 168(3): 723-729.
[18]	HechtV, Vielle-Calzada J P, HartogM V, et al. The Arabidopsis somatic embryogenesis receptor kinase1 gene is expressed in developing ovules and embryos and enhances embryogenic competence in culture[J]. Plant Physiol, 2001. 127: 803-816.
[19]	陈小飞, 萧浪涛, 鲁旭东, 等. 体细胞胚胎发生相关类受体蛋白激酶基因(SERK)的研究进展[J]. 植物生理学通讯, 2005,41(5): 570-577.
[20]	石雅丽. 棉花体细胞胚发生受体类激酶基因的克隆与功能分析[D]. 北京:中国农业科学院研究生院, 2011.
[21]	Zhao D Z, Wang G F, Speal B,et al. The EXCESS MICROSPOROCYTES1 gene encodes a putative leucine-rich repeat receptor protein kinase that controls somatic and reproductive cell fates in the Arabidopsis anther[J].Genes Dev, 2002,16(15): 2021-2031.
[22]	Kemmerling B, Schwedt A, Rodriguez P, et al. Isolation and characterization of four somatic embryogenesisreceptor-like kinase (RhSERK) genes from miniature potted rose (Rosa hybrida cv. Linda)[J]. Plant Cell Tiss Organ Cult, 2010,101(3): 331-338.
[23]	He K, Gou X P, Yuan T, et al. BAK1 and BKK1 regulate brassinosteroid-dependent growth and brassinosteroid-independent cell-death pathways[J]. Curr Biol, 2007,17(13): 1109-1115.
[24]	Emidio A, Gianpiero M, Gianni Ba, et al. Isolation of candidate genes for apomixis in Poa pratensis L[J]. Plant Molecular Bioligy, 2004,56: 879-894.

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Isolation and Bioinformatics Analysis of Apomicxis SERK1 Genes in Malus

1. College of Forestry, Shenyang Agricultural University, Shenyang 110866, Liaoning, China

2. College of Horticulture, Shenyang Agricultural University, Shenyang 110866, Liaoning, China

3. Forestry Bureau of Huanren County, Benxi 117201, Liaoning, China

Received Date: 2015-06-26

Abstract: The plants Malus hupehensis var. pingyiensis Jiang (Pingyi Tiancha) and a hybrid strain 33# were employed as the experimental materials. By PCR techniques and primers which were designed from the CDS sequences of apple genome, the full-length cDNA sequences of SERK homologous genes were cloned, which were named as MhSERK1 and MhdSERK1 (GenBank accession No. JQ231273 and JQ231272), and then SERKs expressions were detected in different tissues and organs of Pingyi Tiancha and the hybrid strain through Real-time quantitative PCR method. The results showed that the length of coding region sequences about MhSERK1 and MhdSERK1 were 1 899 bp and 1 881 bp, which respectively encoded 632 and 626 amino acids. Compared with other plants, the amino acid sequence homology of SERK1 was 80% or more, especially with Longan grape (Vitaceae), which could reach 92.56%, and it also had a high homology with the model plant Arabidopsis thaliana and tobacco. The results of real-time quantitative PCR showed that the expression levels of SERK1 gene were different among the different tissues and organs of Pingyi Tiancha and hybrid strain, which was high expression levels in the ovary, very low expression levels in vegetative tissues, and the highest expression levels in the ovary of the flower bud of Pingyi Tiancha, indicating that SERK1 gene played an important role in the reproductive and developmental processes of Pingyi Tiancha and hybrid strain.

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Reference (24)

[1]	束怀瑞.苹果学[M].北京:中国农业出版社,1999,210-235.
[2]	李光杰. 平邑甜茶SnRK1基因的克隆、表达及其功能的研究[D]. 泰安:山东农业大学, 2009.
[3]	史兴征. 平邑甜茶GLB1基因的克隆、表达及其功能的研究[D].泰安:山东农业大学, 2010.
[4]	张丽杰,苹果属无融合生殖相关SERK基因克隆与表达分析及遗传转化的研究[D]. 沈阳:沈阳农业大学, 2013.
[5]	Schmidt H. Contribution on the Breeding of Apomictic Apple Stocks 4. On the Inheritance of Apomixis[J]. Z Pflanzenzuchtg. 1977,78:312.
[6]	林盛华, 刘捍中, 蒲富慎. 苹果属植物不同倍性的杂种后代染色体数目观察[J]. 中国果树, 1989,(4): 15-16.
[7]	王颖, 商月惠, 王玉霞, 等.平邑甜茶与扎矮山定子杂交后代的胚胎发育特征研究[J]. 园艺学报, 2008,35(8):1093-1100.
[8]	王颖. 平邑甜茶(Malus hupehensis var.pingyiensis)的杂交后代生殖特性研究和花期子房抑制性消减文库构建[D]. 沈阳:沈阳农业大学, 2009.
[9]	林庆光, 崔百明, 彭明. SERK 基因家族的研究进展[J]. 遗传, 2007, 29(6): 681-687.
[10]	Schmidt E D, Guzzo F, Toonen M A, et al. A leucine-rich repeat containing receptor-like kinase marks somatic plant cells competent to form embryos[J]. Development, 1997, 124(10): 2049-2062.
[11]	Albrecht C, Russinova E, Hecht V, et al. The Arabidopsis thaliana SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASES1 and 2 control male sporogenesis[J]. Plant Cell, 2005,17(12): 3337-3349.
[12]	Colcombet J, Boisson-Dernier A, Ros-Palau R, et al. Arabidopsis SOMATIC EMBRYOGENESIS RECEPTOR KINASES1 and 2 are essential for tapetum development and microspore maturation[J]. Plant Cell, 2005, 17(12): 3350-3361.
[13]	Hecht V, Vielle-Calzada J P, Hartog M V, et al. The Arabidopsis somatic embryogenesis receptor kinase 1 gene is expressed in developing ovules and embryos and enhances embryogenic competence in culture[J]. Plant Physiol, 2001, 127(3): 803-816.
[14]	石雅丽, 张锐, 林芹, 等. 植物体细胞胚胎发生受体类蛋白激酶的生物学功能[J]. 遗传, 2012,34(5): 551-559.
[15]	邹冬梅,刘月学,张志宏,等. 草莓AP1同源基因的克隆与表达分析[J]. 中国农业科学,2012, 45(10):1972-1981.
[16]	Jun Ma, Yehua He, Chenghou Wu, et al. Cloning and molecular characterization of a SERK gene transcriptionally induced during somatic embryogenesis in Ananas comosus cv. Shenwan[J]. Plant Mol Biol Rep,2012, 30: 195-203.
[17]	Santos M D O, Romano E, Yotoko K S C, et al. Characterisation of the cacao somatic embryogenesis receptor-like kinase (SERK) gene expressed during somatic embryogenesis[J]. Plant Sci, 2005, 168(3): 723-729.
[18]	HechtV, Vielle-Calzada J P, HartogM V, et al. The Arabidopsis somatic embryogenesis receptor kinase1 gene is expressed in developing ovules and embryos and enhances embryogenic competence in culture[J]. Plant Physiol, 2001. 127: 803-816.
[19]	陈小飞, 萧浪涛, 鲁旭东, 等. 体细胞胚胎发生相关类受体蛋白激酶基因(SERK)的研究进展[J]. 植物生理学通讯, 2005,41(5): 570-577.
[20]	石雅丽. 棉花体细胞胚发生受体类激酶基因的克隆与功能分析[D]. 北京:中国农业科学院研究生院, 2011.
[21]	Zhao D Z, Wang G F, Speal B,et al. The EXCESS MICROSPOROCYTES1 gene encodes a putative leucine-rich repeat receptor protein kinase that controls somatic and reproductive cell fates in the Arabidopsis anther[J].Genes Dev, 2002,16(15): 2021-2031.
[22]	Kemmerling B, Schwedt A, Rodriguez P, et al. Isolation and characterization of four somatic embryogenesisreceptor-like kinase (RhSERK) genes from miniature potted rose (Rosa hybrida cv. Linda)[J]. Plant Cell Tiss Organ Cult, 2010,101(3): 331-338.
[23]	He K, Gou X P, Yuan T, et al. BAK1 and BKK1 regulate brassinosteroid-dependent growth and brassinosteroid-independent cell-death pathways[J]. Curr Biol, 2007,17(13): 1109-1115.
[24]	Emidio A, Gianpiero M, Gianni Ba, et al. Isolation of candidate genes for apomixis in Poa pratensis L[J]. Plant Molecular Bioligy, 2004,56: 879-894.

Isolation and Bioinformatics Analysis of Apomicxis SERK1 Genes in Malus

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通讯作者: 陈斌, bchen63@163.com

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Isolation and Bioinformatics Analysis of Apomicxis SERK1 Genes in Malus

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