Cloning and Expression Pattern Analysis of DfDXR Gene in Dryopteris fragrans

doi:10.7668/hbnxb.20193254

Abstract

Abstract:

In order to explore whether 1-deoxygen-D-xylose-5-phosphate reductisomerase(DXR)participates in the environmental resilience of Dryopteris fragrans.It used polymerase chain reaction(Polymerase Chain Reaction,PCR)methods to clone and identify Dryopteris fragrans DfDXR gene,the protein sequences were analyzed by bioinformatics through online prediction website,the relative expression levels of DfDXR gene under different chemical substances and stress were studied by Real-time fluorescence quantitative PCR(qRT-PCR).The experiment further explored how DfDXR gene responded to the stress-resistance process of Dryopteris fragrans,and laid a foundation for studying its mechanism.Experimental results:The full-length CDS sequence of DfDXR gene 1 434 bp was successfully cloned,encoding 477 amino acids,and the secondary structure of DXR protein was alpha-beta.Multi-sequence alignment and phylogenetic tree analysis of the proteins suggested that DfDXR was closely related to Adiantum capillus-Veneris AcDXR,but far related to DXR protein of apple and peach.Motif analysis showed that the protein contained PLN02696 domain,which was the conserved domain of DXR protein.Subcellular localization predicted that DXR protein was located in chloroplast.The analysis of qRT-PCR data showed that the relative expression of DfDXR showed an upward trend after treatment with methyl 3-oxo-2-(pent-2-enyl)cyclamate(MeJA)and polyethyleneglycol(PEG),and reached the highest level at 1,12 h respectively.Under NaCl treatment,there was a "down—up—down" trend,but the expression levels under each treatment time were lower than those in the control group.The relative expression of DfDXR also changed obviously under the treatment of salicylic acid(SA),abscisic acid(ABA),ethephon(ETH),high teperature(HT)and low temperature(LT).DfDXR plays a regulatory role in the response of Dryopteris fragrans to different abiotic stresses,and its response time to different chemical substances and stress is different.

Key words: Dryopteris fragrans, DfDXR genes, Abiotic stress, Terpenoids, Function verification

MA Qian, YUAN Qiang, QIU Xiaojie, SONG Chunhua, CHANG Ying. Cloning and Expression Pattern Analysis of DfDXR Gene in Dryopteris fragrans[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(1): 102-109. doi: 10.7668/hbnxb.20193254.

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References 29

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生物信息学分析 Bioinformatics analysis	网站 Website
理化性质 Physicochemical property	http://web.expasy.org/protparam/
亚细胞定位Subcellular localization	http://www.csbio.sjtu.edu.cn/bioinf/plant-multi/
二级结构 Secondary structure	https://npsa-prabi.ibcp.fr/cgi-bin/npsa_automat.pl?page=/NPSA/npsa_sopma.html
三级结构 Tertiary structure	http://swissmodel.expasy.org/
磷酸化位点 Phosphorylation site	http://www.cbs.dtu.dk/services/NetPhos
保守序列分析 Motif analysis	http://meme-suite.org/index.html

生物信息学分析 Bioinformatics analysis	网站 Website
理化性质 Physicochemical property	http://web.expasy.org/protparam/
亚细胞定位Subcellular localization	http://www.csbio.sjtu.edu.cn/bioinf/plant-multi/
二级结构 Secondary structure	https://npsa-prabi.ibcp.fr/cgi-bin/npsa_automat.pl?page=/NPSA/npsa_sopma.html
三级结构 Tertiary structure	http://swissmodel.expasy.org/
磷酸化位点 Phosphorylation site	http://www.cbs.dtu.dk/services/NetPhos
保守序列分析 Motif analysis	http://meme-suite.org/index.html

Motif	长度/aa Length	序列(5'—3') Sequence	名称 Description
1	50	HPQSIIHSMIETQDSSVLAQLGWPDMRLPILYTMSWPD RVPCSEITWPRL	DXP还原异构酶C端结构域
2	50	FRDWPVEKLKEVKVADALKHPNWNMGKKITVDSATL FNKGLEVIEAHYLF	1-脱氧-D-木酮糖-5-磷酸还原异构酶C端结构域(缩短)
3	50	YPSMDLAYAAGRAGGTMTGVLSAANEKAVEMFJDEKI SYLDIFKVVELTC	DXP还原异构酶C端结构域
4	50	GKDIALANKETLIAGGPFVLPLANKHNVKILPADSEHSA IFQCIQGLPEG	1-脱氧-D-木酮糖-5-磷酸还原异构酶C端结构域(缩短)
5	50	GPKPISIVGSTGSIGTQTLDIVAENPDKFRVVALAAGSNV TLLADQVKRF	1-脱氧-D-木酮糖-5-磷酸还原异构酶(缩短)
6	50	EALADLDEKPEIIPGEQGVIEVARHPDAVTVVTGIVGCA GLKPTVAAIEA	1-脱氧-D-木酮糖-5-磷酸还原异构酶(缩短)
7	29	KHRNELVSSPSLEEIVHYDLWAREYAANV	未知
8	21	AQPPPPAWPGRAVPETGRKSW	未知
9	15	DLCKLGSLTFKAPDN	未知
10	15	KPQLVAVRNESLVNE	未知

Motif	长度/aa Length	序列(5'—3') Sequence	名称 Description
1	50	HPQSIIHSMIETQDSSVLAQLGWPDMRLPILYTMSWPD RVPCSEITWPRL	DXP还原异构酶C端结构域
2	50	FRDWPVEKLKEVKVADALKHPNWNMGKKITVDSATL FNKGLEVIEAHYLF	1-脱氧-D-木酮糖-5-磷酸还原异构酶C端结构域(缩短)
3	50	YPSMDLAYAAGRAGGTMTGVLSAANEKAVEMFJDEKI SYLDIFKVVELTC	DXP还原异构酶C端结构域
4	50	GKDIALANKETLIAGGPFVLPLANKHNVKILPADSEHSA IFQCIQGLPEG	1-脱氧-D-木酮糖-5-磷酸还原异构酶C端结构域(缩短)
5	50	GPKPISIVGSTGSIGTQTLDIVAENPDKFRVVALAAGSNV TLLADQVKRF	1-脱氧-D-木酮糖-5-磷酸还原异构酶(缩短)
6	50	EALADLDEKPEIIPGEQGVIEVARHPDAVTVVTGIVGCA GLKPTVAAIEA	1-脱氧-D-木酮糖-5-磷酸还原异构酶(缩短)
7	29	KHRNELVSSPSLEEIVHYDLWAREYAANV	未知
8	21	AQPPPPAWPGRAVPETGRKSW	未知
9	15	DLCKLGSLTFKAPDN	未知
10	15	KPQLVAVRNESLVNE	未知

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