Plant stress－resistant gene cloning for Na and Cd using an INVSc1 yeast

SONG Hong－gai; JIANG Jing; QIAO Gui－rong; YANG Ye; ZHOU Jing; PAN Luan－yin; ZHUO Ren－ying

doi:10.11833/j.issn.2095-0756.2010.06.014

Volume 27 Issue 6

Dec. 2010

Turn off MathJax

Article Contents

Article Navigation > Journal of Zhejiang A&F University > 2010 > 27(6): 890-895

SONG Hong－gai, JIANG Jing, QIAO Gui－rong, YANG Ye, ZHOU Jing, PAN Luan－yin, ZHUO Ren－ying. Plant stress－resistant gene cloning for Na and Cd using an INVSc1 yeast[J]. Journal of Zhejiang A&F University, 2010, 27(6): 890-895. doi: 10.11833/j.issn.2095-0756.2010.06.014

Citation:

SONG Hong－gai, JIANG Jing, QIAO Gui－rong, YANG Ye, ZHOU Jing, PAN Luan－yin, ZHUO Ren－ying. Plant stress－resistant gene cloning for Na and Cd using an INVSc1 yeast[J]. Journal of Zhejiang A&F University, 2010, 27(6): 890-895. doi: 10.11833/j.issn.2095-0756.2010.06.014

Plant stress－resistant gene cloning for Na and Cd using an INVSc1 yeast

doi: 10.11833/j.issn.2095-0756.2010.06.014

1.
Biotechnology Research Center/Hubei Province Key Laboratory of Natural Products Research and Development， China Three Gorges University，Yichang 443002，Hubei，China;
2.
The Research Institute of Subtropical Forestry，The Chinese Academy of Forestry，Fuyang 311400，Zhejiang，China

More Information

Corresponding author: ZHUO Ren－ying
Publish Date: 2010-12-20

Abstract

To provide a basis for studying a new stress－related gene of Salix matsudana and Sedum alfredii，the salt－stress induced full－length cDNA Library of Salix matsudana and the cadmium－stress induced，full－length cDNA Library of Sedum alfredii were constructed using an improved switching mechanism at 5 end of RNA transcript （SMART） method. The full－length cDNA fragment was linked to the yeast vector pYES2.1，and then the linked product was transferred onto the yeast INVSc1. Concentrations of NaCl and cadmium stress were utilized to construct yeast transformants library. Growth performance of yeast recombinants and their control with an empty vector were tested under the control（0），high salinity （1 molL-1 NaCl），or high cadmium（150 molL-1 cadmium） conditions. Also，a bioinformatics analysis and real－time polymerase chain reaction （PCR） were conducted. Results for the yeast transformants library revealed two positive transformants with a tolerance of 1.709 molL-1 NaCl and two positive transformants with a tolerance of 0.123 mmolL-1 cadmium. Growth rates of the recombinants with cDNA protein under the non－stress condition indicated that the expression of cDNA was not deleterious to yeast growth. Compared to the control，yeast cells expressing cDNA showed a shorter lag period when transferred to a medium containing high salinity and high cadmium. The results of a bioinformatics analysis and real－time PCR showed that the No. 6 salt－tolerant transformant gene was a salt－resistant related gene. This study showed that the expression of cDNA could confer salt and cadmium tolerance in yeast and provided an important tool for establishing an efficient system of stress－resistant gene cloning using the yeast INVSc1.［Ch，9 fig. 12 ref.］
- forest tree breeding,
- Salix matsudana,
- Sedum alfredii,
- cDNA library,
- yeast,
- salt tolerance,
- cadmium tolerance,
- grouth curves

Relative Article

[1]	HUANG Qingchen, LAI Jianxin, HUANG Lichao, LU Mengzhu. Effects of peroxidase gene PagPRX19 on salt tolerance of poplar ‘84K’ . Journal of Zhejiang A&F University, 2022, 39(6): 1163-1172. doi: 10.11833/j.issn.2095-0756.20220387
[2]	LI Dandan, LIN Rong, LI Xinguo, ZHENG Yueping. Functional analysis of AtJAR1 gene in salt tolerance of Arabidopsis thaliana . Journal of Zhejiang A&F University, 2022, 39(5): 998-1009. doi: 10.11833/j.issn.2095-0756.20210742
[3]	LIU Bo, CHEN Ji, MA Jiawei, FANG Xianzhi, ZHAO Keli, LIU Dan, YE Zhengqian. Effects of AC electric field and soil moisture on phytoremediation of cadmium contaminated soil by mixed planting of Salix discolor and Sedum alfredii . Journal of Zhejiang A&F University, 2021, 38(6): 1238-1244. doi: 10.11833/j.issn.2095-0756.20200725
[4]	LIU Liping, DAI Fengbin, ZHANG Chong, TIAN Ju, CHEN Jinhuan. Cloning and expression analysis of the SKOR gene for an outward-rectifying K⁺ channel in Lycium ruthenicum . Journal of Zhejiang A&F University, 2018, 35(1): 104-111. doi: 10.11833/j.issn.2095-0756.2018.01.014
[5]	CHEN Wen, WANG Jing, WU Haiping, ZHANG Ling, YE Zhengqian. Salt tolerance of 5 typical landscape plant species in Zhoushan Islands . Journal of Zhejiang A&F University, 2017, 34(2): 283-293. doi: 10.11833/j.issn.2095-0756.2017.02.012
[6]	LIU Yu, XU Huanwen, LI Zhixin, JIANG Jing, PENG Rusheng, SHANG Fuqiang, ZOU Jianjun, TENG Wenhua, LIU Guifeng. Growth performance and adaptability of Betula platyphylla offspring in the period of young forest . Journal of Zhejiang A&F University, 2015, 32(6): 853-860. doi: 10.11833/j.issn.2095-0756.2015.06.005
[7]	ZHAO Ting, HAN Xiaojiao, LIU Mingying, QIAO Guirong, JIANG Jing, JIANG Yancheng, ZHUO Renying. Isolation and expression of a cadmium-resistant gene （SaFer） from Sedum alfredii . Journal of Zhejiang A&F University, 2015, 32(1): 25-32. doi: 10.11833/j.issn.2095-0756.2015.01.004
[8]	HU Yangyong, MA Jiawei, YE Zhengqian, LIU Dan, ZHAO Keli. Research progress on using Sedum alfredii for remediation of heavy metal-contaminated soil . Journal of Zhejiang A&F University, 2014, 31(1): 136-144. doi: 10.11833/j.issn.2095-0756.2014.01.021
[9]	LI Zhen, LIU Mingying, HAN Xiaojiao, QIAO Guirong, JIANG Jing, XING Shiyan, ZHUO Renying. Characterization of a light-harvesting chlorophyll a/b binding protein (LHCB) gene, SaLhcb2, in Sedum alfredii . Journal of Zhejiang A&F University, 2014, 31(6): 838-846. doi: 10.11833/j.issn.2095-0756.2014.06.003
[10]	ZHANG Zhen, ZHANG Han-guo, ZHANG Lei, ZHU Hang-yong, LI Xue-feng. Diversity analysis with RAPD for natural and breeding populations of Larix gmelinii . Journal of Zhejiang A&F University, 2012, 29(1): 130-136. doi: 10.11833/j.issn.2095-0756.2012.01.022
[11]	HUANG Yin-zhi, ZHOU Qin, HUANG You-jun, ZENG Yan-ru. Construction of a fat metabolism-related cDNA library in Carya cathayensis . Journal of Zhejiang A&F University, 2011, 28(1): 80-85. doi: 10.11833/j.issn.2095-0756.2011.01.013
[12]	ZHANG Xiao-guo, QIAO Gui-rong, LI Cui-yun, LIU Ming-ying, JIANG Jing, LI Hai-ying, QIU Wen-min, ZHUO Ren-ying. A two-dimensional electrophoresis system of Salix matsudana root proteomics . Journal of Zhejiang A&F University, 2011, 28(4): 653-661. doi: 10.11833/j.issn.2095-0756.2011.04.021
[13]	ZHANG Yuan－yuan, DOU Chun－ying, YAO Fang1, YE Zheng－qian. Nitrogen application to enhance zinc and cadmium uptake by the hyperaccumulator Sedum alfredii . Journal of Zhejiang A&F University, 2010, 27(6): 831-838. doi: 10.11833/j.issn.2095-0756.2010.06.005
[14]	WANG Hong－ling, ZHANG Jue, JIAO Zhong－yi, SHI De－zong. Establishing and optimizing the SRAP－PCR system for Salix matsudana . Journal of Zhejiang A&F University, 2010, 27(6): 945-949. doi: 10.11833/j.issn.2095-0756.2010.06.024
[15]	XU Ai-chun, CHEN Yi-tai, CHEN Qing-hong, WANG Shu-fen. Cd tolerance and leaf physiology changes in ten clones of Salix matsudana with Cd stress . Journal of Zhejiang A&F University, 2009, 26(5): 674-681.
[16]	YANG Ye, JIANG Jing, QIAO Gui-rong, ZHOU Jing, CHEN Yin, HE Zheng-quan, LI Hai-ying, ZHUO Ren-ying. Expressed salt stress induced full length-cDNA library of Salix matsudana using yeast Invsc1 . Journal of Zhejiang A&F University, 2009, 26(4): 473-478.
[17]	Zhang Liqin, Zheng Yong ping, Luo Shiyuan, Hu Jiagong.. Salt Tolerance of Calli of Populus Clones and Pinus elliottii in Tissue Cultures . Journal of Zhejiang A&F University, 1997, 14(1): 16-21.
[18]	Wan Fuxu, Hu HaibO, Zhang Jinchi, Jiang Furong, Sun Wenkang. Growth Characteristies of Populus Constituting the Agricultural Protcetion Forest in the Northern Area of the Huaihe River. . Journal of Zhejiang A&F University, 1996, 13(3): 280-285.
[19]	Zhang Liqin, Zheng YongPing, Jin Peiying. selection of Salt Tolerance poplar Regenerated plant-lets in Tissues Culture. . Journal of Zhejiang A&F University, 1996, 13(4): 397-404.
[20]	Luo Fuyu, Fang Yanfu, Li Longchang. Study on Growth Model of Man-made Forest of Chinese Fir in Suichang County . Journal of Zhejiang A&F University, 1993, 10(1): 49-54.

References
Proportional views

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

Get Citation

PDF

XML

Article views(5376) PDF downloads(1137) Cited by()

Proportional views

Plant stress－resistant gene cloning for Na and Cd using an INVSc1 yeast

1. Biotechnology Research Center/Hubei Province Key Laboratory of Natural Products Research and Development， China Three Gorges University，Yichang 443002，Hubei，China;

2. The Research Institute of Subtropical Forestry，The Chinese Academy of Forestry，Fuyang 311400，Zhejiang，China

Corresponding author: ZHUO Ren－ying

Publish Date: 2010-12-20

Key words:

Abstract: To provide a basis for studying a new stress－related gene of Salix matsudana and Sedum alfredii，the salt－stress induced full－length cDNA Library of Salix matsudana and the cadmium－stress induced，full－length cDNA Library of Sedum alfredii were constructed using an improved switching mechanism at 5 end of RNA transcript （SMART） method. The full－length cDNA fragment was linked to the yeast vector pYES2.1，and then the linked product was transferred onto the yeast INVSc1. Concentrations of NaCl and cadmium stress were utilized to construct yeast transformants library. Growth performance of yeast recombinants and their control with an empty vector were tested under the control（0），high salinity （1 molL-1 NaCl），or high cadmium（150 molL-1 cadmium） conditions. Also，a bioinformatics analysis and real－time polymerase chain reaction （PCR） were conducted. Results for the yeast transformants library revealed two positive transformants with a tolerance of 1.709 molL-1 NaCl and two positive transformants with a tolerance of 0.123 mmolL-1 cadmium. Growth rates of the recombinants with cDNA protein under the non－stress condition indicated that the expression of cDNA was not deleterious to yeast growth. Compared to the control，yeast cells expressing cDNA showed a shorter lag period when transferred to a medium containing high salinity and high cadmium. The results of a bioinformatics analysis and real－time PCR showed that the No. 6 salt－tolerant transformant gene was a salt－resistant related gene. This study showed that the expression of cDNA could confer salt and cadmium tolerance in yeast and provided an important tool for establishing an efficient system of stress－resistant gene cloning using the yeast INVSc1.［Ch，9 fig. 12 ref.］

Plant stress－resistant gene cloning for Na and Cd using an INVSc1 yeast

doi: 10.11833/j.issn.2095-0756.2010.06.014

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Related

Proportional views