Cloning of the <em>PmPIP</em>1 gene from Pinus massoniana and its expression with drought stress

CAI Qiong; DING Guijie; WEN Xiaopeng

doi:10.11833/j.issn.2095-0756.2016.02.002

Volume 33 Issue 2

Mar. 2016

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CAI Qiong, DING Guijie, WEN Xiaopeng. Cloning of the PmPIP1 gene from Pinus massoniana and its expression with drought stress[J]. Journal of Zhejiang A&F University, 2016, 33(2): 191-200. doi: 10.11833/j.issn.2095-0756.2016.02.002

Citation:

CAI Qiong, DING Guijie, WEN Xiaopeng. Cloning of the PmPIP1 gene from Pinus massoniana and its expression with drought stress[J]. Journal of Zhejiang A&F University, 2016, 33(2): 191-200. doi: 10.11833/j.issn.2095-0756.2016.02.002

Cloning of the PmPIP1 gene from Pinus massoniana and its expression with drought stress

doi: 10.11833/j.issn.2095-0756.2016.02.002

CAI Qiong^1,2,
DING Guijie^{1,2
,
,},
WEN Xiaopeng³

1.
Institute of Forest Resources and Environment, Guiyang 550025, Guizhou, China;
2.
College of Forestry, Guizhou University, Guiyang 550025, Guizhou, China;
3.
Guizhou Key Laboratory of Agricultural Bioengineering, Guizhou University, Guiyang 550025, Guizhou, China

Funds:

国家自然科学基金资助项目（31260183）;十二五国家科技支撑计划项目（2015BAD09B0102）;国家高技术研究发展计划（863计划）项目（2011AA10020301）;贵州省重大专项（黔科合重大专项字［2012］6001号）;贵州省人才基地建设项目（黔人领发［2009］9号）

Received Date: 2015-04-10
Rev Recd Date: 2015-05-18
Publish Date: 2016-04-20

Abstract

The aquaporin （AQP） gene plays an important role in plants adapting to abiotic stresses. To predict the AQP gene function and provide basal data for mechanism of Pinus massonianas drought resistance, the sequence characteristics of the AQP gene from P. massoniana were analyzed and its expression profiling was studied after drought-stress treatment. The AQP gene was cloned using reverse transcription-polymerase chain reaction （RT-PCR） and rapid-amplification of complementary deoxyribonucleic acid （cDNA） ends （RACE）. The expression of the AQP gene was then performed using quantitative reverse transcriptase-polymerase chain reaction （RT-qPCR）. The full-length cDNA of the AQP gene from P. massoniana, designated PmPIP1 with a registered number in GenBank KF582038, was obtained. Results of the sequence analysis showed that the size of PmPIP1 was 1 301 bp, containing an 867 bp open reading frame that encoded 288 amino acid residues with 30.86 kDa molecular weight and an 8.48 isoelectric point, a 99 bp 5 terminal untranslated regions （UTR）, and a 335 bp 3 terminal UTR. The PmPIP1 3D structure had a strong similarity to Spinacia oleracea （2b5fA）. PmPIP1 exhibited a typical structure with six transmembrane domains, and had the consensus sequence HINPAVTFG of membrane intrinsic protein （MIP） family and two highly conserved peptides Asn-Pro-Ala （NPA）. The evolutionary analysis revealed that PmPIP1 shared a 95% identity with Picea abies and belonged to PIPs. The PmPIP1 expression patterns with drought conditions showed that drought did induce PmPIP1. In conclusion, cloning of the PmPIP1 gene from P. massoniana enriched the plant aquaporin gene database, and the RT-qPCR analysis indicated that the PmPIP1 gene may be involved in the response related to drought stress. ［Ch, 8 fig. 1 tab. 39 ref.］
- botany,
- Pinus massoniana,
- aquaporin,
- cloning,
- expression

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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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Cloning of the PmPIP1 gene from Pinus massoniana and its expression with drought stress

1. Institute of Forest Resources and Environment, Guiyang 550025, Guizhou, China;

2. College of Forestry, Guizhou University, Guiyang 550025, Guizhou, China;

3. Guizhou Key Laboratory of Agricultural Bioengineering, Guizhou University, Guiyang 550025, Guizhou, China

Funds:

Received Date: 2015-04-10

Rev Recd Date: 2015-05-18

Publish Date: 2016-04-20

Key words:

Abstract: The aquaporin （AQP） gene plays an important role in plants adapting to abiotic stresses. To predict the AQP gene function and provide basal data for mechanism of Pinus massonianas drought resistance, the sequence characteristics of the AQP gene from P. massoniana were analyzed and its expression profiling was studied after drought-stress treatment. The AQP gene was cloned using reverse transcription-polymerase chain reaction （RT-PCR） and rapid-amplification of complementary deoxyribonucleic acid （cDNA） ends （RACE）. The expression of the AQP gene was then performed using quantitative reverse transcriptase-polymerase chain reaction （RT-qPCR）. The full-length cDNA of the AQP gene from P. massoniana, designated PmPIP1 with a registered number in GenBank KF582038, was obtained. Results of the sequence analysis showed that the size of PmPIP1 was 1 301 bp, containing an 867 bp open reading frame that encoded 288 amino acid residues with 30.86 kDa molecular weight and an 8.48 isoelectric point, a 99 bp 5 terminal untranslated regions （UTR）, and a 335 bp 3 terminal UTR. The PmPIP1 3D structure had a strong similarity to Spinacia oleracea （2b5fA）. PmPIP1 exhibited a typical structure with six transmembrane domains, and had the consensus sequence HINPAVTFG of membrane intrinsic protein （MIP） family and two highly conserved peptides Asn-Pro-Ala （NPA）. The evolutionary analysis revealed that PmPIP1 shared a 95% identity with Picea abies and belonged to PIPs. The PmPIP1 expression patterns with drought conditions showed that drought did induce PmPIP1. In conclusion, cloning of the PmPIP1 gene from P. massoniana enriched the plant aquaporin gene database, and the RT-qPCR analysis indicated that the PmPIP1 gene may be involved in the response related to drought stress. ［Ch, 8 fig. 1 tab. 39 ref.］

Cloning of the PmPIP1 gene from Pinus massoniana and its expression with drought stress

doi: 10.11833/j.issn.2095-0756.2016.02.002

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

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Proportional views

Cloning of the PmPIP1 gene from Pinus massoniana and its expression with drought stress

doi: 10.11833/j.issn.2095-0756.2016.02.002

1. Institute of Forest Resources and Environment, Guiyang 550025, Guizhou, China;

2. College of Forestry, Guizhou University, Guiyang 550025, Guizhou, China;

3. Guizhou Key Laboratory of Agricultural Bioengineering, Guizhou University, Guiyang 550025, Guizhou, China

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Cloning of the PmPIP1 gene from Pinus massoniana and its expression with drought stress

doi: 10.11833/j.issn.2095-0756.2016.02.002

Abstract

Relative Article

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Related

Proportional views

Cloning of the PmPIP1 gene from Pinus massoniana and its expression with drought stress

doi: 10.11833/j.issn.2095-0756.2016.02.002

1. Institute of Forest Resources and Environment, Guiyang 550025, Guizhou, China; 2. College of Forestry, Guizhou University, Guiyang 550025, Guizhou, China; 3. Guizhou Key Laboratory of Agricultural Bioengineering, Guizhou University, Guiyang 550025, Guizhou, China

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1. Institute of Forest Resources and Environment, Guiyang 550025, Guizhou, China;

2. College of Forestry, Guizhou University, Guiyang 550025, Guizhou, China;

3. Guizhou Key Laboratory of Agricultural Bioengineering, Guizhou University, Guiyang 550025, Guizhou, China