Reference Gene Selection for qRT-PCR Normalization in Iris germanica L.

Quantitative real-time PCR(qPCR)is an effective and widely used method to analyze expression patterns of target genes.Selection of stable reference genes is a prerequisite for accurate normalization of target gene expression by qRT-PCR.In Iris germanica L.,no studies have yet been published regarding the evaluation of potential reference genes.In this study,nine candidate reference genes were assessed at different flower developmental stages and in different tissues by four different algorithms(GeNorm,NormFinder,BestKeeper,and RefFinder).The results revealed that ACT11(Actin 11)and EF1α(Elongation factor 1 alpha)were the most stable reference genes in different tissues,whereas TUA(Tubulin alpha)and UBC9(Ubiquitin-protein ligase 9)were the most stable ones in different flower developmental stages.UBC9 and ACT11 were the most stable reference genes in all of the tested samples,while the SAMDC(S-Adenosylmethionine decarboxylase)showed the least stability.Finally,to validate the suitability of the selected reference genes,the relative expression level of IgTPS(beta-caryophyllene synthase)was assessed and highlighted the importance of suitable reference gene selection.This work constitutes the first systematic evaluation of potential reference genes in I.germanica and provides guidelines for future research on gene function and molecular mechanisms on I.germanica and related species.

FRANS HALS德国鸢尾组织培养及植株再生研究

以德国鸢尾(Iris germanica.L)FRANS HALS叶片、球茎作为外植体,利用正交试验设计研究探讨不同激素及不同浓度组合对鸢尾愈伤组织的诱导、不定芽分化以及试管苗生根的影响,为鸢尾快速繁殖、种质资源的保存及利用提供最优的技术方法。结果表明,不同外植体灭菌时间也不同,叶片为3%次氯酸钠3 min,球茎为3%次氯酸钠5 min;鸢尾组培快繁的最适合外植体为球茎;鸢尾愈伤组织诱导和芽分化的最适培养基配方为MS+NAA 0.10 mg/L+6-BA 1.00 mg/L,生根培养基为MS+1\2MS+NAA 0.10 mg/L,试管苗移栽基质采用泥炭∶珍珠岩=8∶2,成活率达90%以上。

基于转录组测序信息的德国鸢尾MADS-box基因家族分析

为探究MADS-box基因家族对德国鸢尾多季花品种花期的影响,本研究基于德国鸢尾(Iris germanica)转录组数据,利用生物信息学方法对MADS-box家族基因进行筛选鉴定,并对其理化性质、功能域及亲/疏水性等方面进行预测分析,且构建德国鸢尾和模式植物拟南芥MADS-box蛋白家族的系统进化树。结果表明,在德国鸢尾中鉴定出39个MADS-box基因家族成员,其编码的氨基酸序列分子量在4704.58~30112.19 k D之间,理论等电点(pI)介于5.01~10.92之间,其中32个表现为碱性蛋白,7个表现为酸性蛋白。大多数MADS-box蛋白定位于细胞核和线粒体基质,少许分布在其他细胞器均不含有信号肽。α-螺旋和无规则卷曲为二级结构的主要原件,所有蛋白均表现为亲水性蛋白。进化树分析结果显示,德国鸢尾MADS-box蛋白可聚为5大类(MIKC,Mα,Mβ,Mγ,Mδ)。本研究可为今后进一步研究德国鸢尾MADS-box蛋白的生物学功能提供一定的参考依据。