南方农业学报2023，Vol.54Issue(12)：3464-3474,11.DOI:10.3969/j.issn.2095-1191.2023.12.002
龙州凤仙花ImLHY和ImC4H基因克隆及表达分析
Cloning and expression analysis of ImLHY and ImC4H genes in Impatiens morsei Hook.f.
摘要
Abstract
[Objective]Cloning and expression analysis of ImLHY and ImC4H genes were carried out to investigate the color difference between variegation and non-variegation areas in vexil and wing petals of Impatiens morsei.Cloning of LHY and C4H genes provided theoretical basis for molecular regulation mechanism of flower variegation,flower color improvement and new variety breeding of Impatiens.[Method]The I.morsei as the material,the colorimetric values,con-tents of total anthocyanins and total flavonoids of variegation and non-variegation areas in vexil and wing petals were de-termined.The ImLHY and ImC4H genes were cloned,and their sequence characteristics,protein structure and phylogeny were analyzed by biological software.Real-time quantitative PCR was used to detect the expression patterns of ImLHY and ImC4H genes at different developmental stages(bud stage,blooming stage and decay stage)and in different colored regions of vexil and wing petals.[Result]The a* value(redness value)and C* value(chroma value)of the variegation were higher than the non-variegation area in vexil and wing petals,and L*(brightness value)of the non-variegation area was is greater than the variegation area;and the b* value(yellowness value)of the variegation area was higher than that of the non-variegation area in vexil petal,while the b* value(yellowness value)of the variegation area was less different from that of the non-variegation area in wing petal.The contents of total anthocyanins and flavonoids in vexil and wing petals were higher than those in non-variegation area.The full-length cDNA sequences of ImLHY and ImC4H genes were 1698bp and 1518bp,encoding 565aa and 505aa,respectively.The GenBank login numbers were No.OR096417 and No.OR096417.ImLHY and ImC4H genes were hydrophilic unstable proteins without transmembrane domains.ImLHY pro-tein had a Myb DNA-binding conserved domain and a SANT domain of the MYB transcription factor family,which was localized in the nucleus;ImC4H protein contained a P450 superfamily domain(PLN02394),and located in the endoplas-mic reticulum The amino acid sequences of the ImLHY and ImC4H proteins had 68%and 94%similarity with Impatiens glandulifera.In phylogenetic evolutionary tree,both of them were clustered together with LHY and C4H of I.glandu-lifera respectively.The two genes were expressed in the three key stages of flower development(bud stage,blooming stage and decay stage)and in the variegation and non-variegation areas of vexil and wing petals,and the expression level was the highest at the blooming stage.At blooming stage,the expression levels of ImLHY and ImC4H genes were higher in variegation area of vexil petals than in non-variegation area,and higher in non-variegation area of the wing petals than in variegation area.[Conclusion]ImLHY and ImC4H genes mainly regulate the formation of variegation in vexil petals during the blooming stage,and mainly regulate the formation of variegation in wing petals.It was speculate that there were different regulatory mechanisms for the formation of variegation in vexil and wing petals of I.morsei.关键词
龙州凤仙花/旗瓣/翼瓣 彩斑/LHY基因/C4H基因/表达分析Key words
Impatiens morsei/variegation/phenotypic assay/LHY gene/C4H gene/expression analysis分类
农业科技引用本文复制引用
赵潞秋,张晓丽,李凡,谭弋,石万磊,黄美娟,黄海泉..龙州凤仙花ImLHY和ImC4H基因克隆及表达分析[J].南方农业学报,2023,54(12):3464-3474,11.基金项目
国家自然科学基金项目(32060364,32060366) （32060364,32060366）
云南省农业联合专项重点项目(202301BD070001-011) （202301BD070001-011）
云南省重大科技专项(202102AE090052) （202102AE090052）
云南省园林植物遗传改良与高效繁育博士生导师团队(503210103) National Natural Science Foundation of China,(32060364,32060366) （503210103）
Key Project of Agricul-tural Joint Project of Yunnan Province(202301BD070001-011) （202301BD070001-011）
Key Science and Technology Project of Yunnan Prov-ince(202102AE090052) （202102AE090052）
A Doctoral Supervisor Team for Genetic Improvement and Efficient Breeding of Garden Plants in Yunnan Province(503210103) （503210103）
