菊花花瓣伸长相关基因CmXTHs 的克隆与功能验证

doi:10.16420/j.issn.0513-353x.2017-0129

摘要/Abstract

摘要：

从杭白菊（Chrysanthemum morifolium‘Hangbaiju’）中克隆了4 个细胞壁松弛和伸展相关的
木葡聚糖内转糖苷酶/水解酶基因，分别命名为CmXTH1、CmXTH2、CmXTH3 和CmXTH4，序列分析表
明其编码的氨基酸序列N 端都含有23 ~ 30 个氨基酸组成的信号肽区域和保守的催化活性位点DE（I/L）
DEFLG 以及紧邻的N（R/A）T 组成的N 端糖基化位点，CmXTH1/2/3 的C 端均含有4 个保守的半胱氨
酸残基，CmXTH1/2/3/4 蛋白的相似度为66.2%。系统进化分析结果表明，CmXTH1/2/3 属于XTH 家族Ⅰ
组，CmXTH4 属于Ⅱ组。实时荧光定量结果表明：（1）菊花CmXTH1/2/3/4 在根、茎、叶和花蕾中均有表
达。CmXTH1 在根中表达量较高，CmXTH2/3 在茎中表达量较高，CmXTH4 在花蕾中表达量最高。（2）
CmXTH1/2/3/4 在花序不同部位表达量有差异，CmXTH1/4 在舌状花中表达量最高，CmXTH2/3 在筒状花
中表达量较高。（3）CmXTH1/2/3/4 在舌状花不同发育阶段表达量差异显著，CmXTH1/2 在盛花期表达量
最高，CmXTH3 在衰老期表达量最高，CmXTH4 在初开期表达量最高。病毒诱导基因沉默结果表明，
CmXTH1/2/3/4 沉默系的花序直径和舌状花长度与对照相比均有所减小，其中CmXTH4 沉默系减小最大，
分别减小了25.67%和10.42%，舌状花花瓣表皮细胞与对照相比明显变小，影响了舌状花花瓣的伸长；
CmXTH2 沉默系的筒状花雄蕊长度和CmXTH4 沉默系的花萼直径分别与对照相比显著减小。这表明菊花
CmXTH1/2/3/4 基因参与了花序的开放，促进了舌状花花瓣的伸长，对于菊花的花序增大具有重要作用。

关键词: 菊花, 花瓣, CmXTH, 基因表达, 基因沉默

Abstract:

Four xyloglucan endotransglucosylase/hydrolase genes that related to cell wall loosen were
cloned from chrysanthemum and named as CmXTH1，CmXTH2，CmXTH3 and CmXTH4. Sequences
analysis showed that the N-terminal regions of deduced amino acid sequences all had signal peptides
composed of 20–30 amino acids，conserved catalytic sites of DE（I/L）DEFLG and N-glycosylation sites composed of N（R/A）T，CmXTH1/2/3 had four cysteine in C-terminal regions，the identity of 4 CmXTH1/2/3/4
was 66.2%. The phylogenetic analysis result suggested that CmXTH1/2/3 belongs to groupⅠof XTH
family，CmXTH4 belongs to groupⅡ. The real-time PCR results showed that：（1）CmXTH1/2/3/4 all had
expression in root，stem and leaf. CmXTH1 was highly expressed in chrysanthemum root. CmXTH2/3 was
expressed more in stem. CmXTH4 was highly expressed in alabastrum.（2）CmXTH1/2/3/4 showed
different expression levels in different parts of inflorescence. CmXTH1/4 was expressed more in ray florets.
CmXTH2/3 was expressed more in tubular florets. （3）CmXTH1/2/3/4 showed different expression levels in
different stages of ray florets development. CmXTH1/2 was highly expressed in ray florets at bloom stage，
CmXTH3 was highly expressed at senescent stage，while CmXTH4 was expressed highest at initiation stage.
The virus induced gene silence results showed that，the inflorescences diameters and ray florets lengths of
CmXTH1/2/3/4-silenced groups all showed decrease compared with control，the CmXTH4-silenced group
decreased most by 25.67% and 10.42% respectively，the sizes of petal epidermal cells decreased
obviously，influenced the extension of ray floret petal. The stamen length of CmXTH2-silenced tubular
florets and the diameter of CmXTH4-silenced sepal all showed significant decrease compared with control.
These results indicated that CmXTH1/2/3/4 participated in the opening of inflorescences，promoted the
extension of ray floret petal ， and they were important for the enlargement of chrysanthemum
inflorescences.

Key words: Chrysanthemum morifolium, petal, CmXTH, gene expression, gene silence

温立柱，孙霞，任红，樊红梅，于媛媛，郭芸珲，郑成淑*. 菊花花瓣伸长相关基因CmXTHs 的克隆与功能验证[J]. 园艺学报, 2017, 44(11): 2150-2162.

WEN Lizhu，SUN Xia，REN Hong，FAN Hongmei，YU Yuanyuan，GUO Yunhui，and ZHENG Chengshu*. Cloning and Functional Verification of Chrysanthemum CmXTHs Genes#br# Related to Petal Elongation[J]. ACTA HORTICULTURAE SINICA, 2017, 44(11): 2150-2162.

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