赛菊芋花叶突变体转录组测序分析

doi:10.11924/j.issn.1000-6850.casb2023-0180

摘要/Abstract

摘要：

为了探索赛菊芋花叶突变体叶色变化的分子机制，以绿叶植株(GL)和花叶变异株(WL)为试材，比较二者叶片表型变化，利用高通量测序技术进行转录组测序分析，结合定量PCR进行验证。结果表明：WL叶片显著变短、变窄，叶脉间为白色，叶色RHS为155A，而GL为137C；叶片SPAD仅为GL的25.85%。转录组测序共获得2098条差异表达基因，WL上调表达1172个；下调表达926个。捕光色素蛋白复合体LHC家族基因、光系统结构蛋白PsaK、psbP等23个已知基因差异表达。KEGG Pathway为光合作用天线蛋白通路和光合电子传递及光合磷酸化通路。本研究从转录水平获得赛菊芋花叶突变体分子水平信息，为深入探索其叶色突变的遗传机制打下了基础。

关键词: 赛菊芋, 花叶突变体, 转录组, 高通量测序, 差异表达基因

Abstract:

To explore the molecular mechanism of leaf color changes in the mosaic mutants of Heliopsis helianthoides, green leaf plants (GL) and mosaic leaf mutants (WL) were used as materials to compare their leaf phenotypic changes, and high-throughput sequencing technology was used for transcriptome sequencing analysis. The further validation was carried out by quantitative PCR technology. The results showed that the WL leaves significantly shortened and narrowed with white color between the leaf veins, leaf color RHS was 155A, and 137C in GL; the SPAD of WL leaves was only 25.85% of GL. A total of 2098 differentially expressed genes were obtained from transcriptome sequencing, among them 1172 were up-regulated and 926 were down-regulated in WL. 23 known genes were expressed differently, including light harvesting pigment protein complex LHC family genes, photosystem structural proteins genes PsaK and psbP. KEGG Pathway was the pathway of photosynthetic antenna protein, photosynthetic electron transfer and photophosphorylation. The molecular information of the mosaic leaf mutants of Heliopsis helianthoides was obtained at the transcription level in this study, laying the foundation for in-depth exploration of the genetic mechanism of leaf color mutation.

Key words: Heliopsis helianthoides, mosaic leaf mutants, transcriptome, high-throughput sequencing, DEGs

秦贺兰, 梁芳, 金莹杉, 李子敬, 张华, 胡雪凡. 赛菊芋花叶突变体转录组测序分析[J]. 中国农学通报, 2023, 39(29): 1-7.

QIN Helan, LIANG Fang, JIN Yingshan, LI Zijing, ZHANG Hua, HU Xuefan. Transcriptomic Analysis of the Mosaic Leaf Mutant in Heliopsis helianthoides[J]. Chinese Agricultural Science Bulletin, 2023, 39(29): 1-7.

图/表 9

参考文献 34

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品种	田间叶片伸展状态		叶长/cm	叶宽/cm	叶厚 /mm	花径/cm	叶色	SPAD值
品种	晴天	阴天	叶长/cm	叶宽/cm	叶厚 /mm	花径/cm	叶色	SPAD值
GL	舒展	舒展	8.34±1.74a	4.7±0.15a	0.29±0.04a	7.6±0.55a	137C	45.96±4.12A
WL	稍内卷	舒展	6.26±0.82b	3.6±0.17b	0.24±0.06a	6.6±0.89a	155A	11.88±0.96B

品种	田间叶片伸展状态		叶长/cm	叶宽/cm	叶厚 /mm	花径/cm	叶色	SPAD值
品种	晴天	阴天	叶长/cm	叶宽/cm	叶厚 /mm	花径/cm	叶色	SPAD值
GL	舒展	舒展	8.34±1.74a	4.7±0.15a	0.29±0.04a	7.6±0.55a	137C	45.96±4.12A
WL	稍内卷	舒展	6.26±0.82b	3.6±0.17b	0.24±0.06a	6.6±0.89a	155A	11.88±0.96B

	All(≥200bp)	≥500bp	≥1000bp	N50	GC/%	Total Length	Max Length	Min Length	Average Length
Transcript	91464	16895	1533	394	41.93	34148011	2583	201	373
Unigene	20710	5489	1044	513	42.02	8684563	2583	201	419

	All(≥200bp)	≥500bp	≥1000bp	N50	GC/%	Total Length	Max Length	Min Length	Average Length
Transcript	91464	16895	1533	394	41.93	34148011	2583	201	373
Unigene	20710	5489	1044	513	42.02	8684563	2583	201	419

功能分类	基因名称	生物学功能	表达情况
叶绿素生物合成	hemA1	编码谷氨酰-tRNA 还原酶，催化叶绿素生物合成	上调
叶绿素生物合成	por	编码原叶绿素酸酯还原酶，催化叶绿素生物合成	上调
叶绿体形成和发育	FTH1	编码铁蛋白重链，维持叶绿体内铁离子的稳态	上调
	CLTC	编码网格蛋白重链，进行叶绿体细胞内的蛋白运输	上调
	RP-S4, rpsD	编码小亚基核糖体蛋白S4，叶绿体核糖体蛋白的构成、翻译	上调
	RP-L9e, RPL9	编码大亚基核糖体蛋白L9e，叶绿体被膜上核糖体的构成	上调
	SOD1	编码Cu-Zn家族超氧化物歧化酶，叶绿体基质、类囊体Cu²⁺、Zn²⁺的结合	上调
	HSPA1/8	编码热激蛋白70B，叶绿体中蛋白的修复	上调
	HSP90B, TRA	编码热激蛋白90B，叶绿体基质中ATP结合、蛋白质折叠	上调
	groES, HSPE1	编码分子伴侣GroES，叶绿体RNA转录、加工、翻译	上调
	VARS, valS	编码缬氨酸-tRNA合成酶，叶绿体氨酰基tRNA活性	上调
	ACSL, fadD	编码长链脂酰CoA合成酶，叶绿体被膜的构成	上调
	ALDO	编码果糖二磷酸化醛缩酶，叶绿体中光合作用参与calvin循环	下调
	HPL	编码脂氢过氧化物裂解酶，叶绿体外膜上催化脂氢过氧化物裂解	下调
	PWYLLO	编码异分支酸合成酶，维持叶绿体膜光合系统Ⅰ的稳定性	下调
	E3.6.3.8	编码Ca²⁺-转运 ATPase，叶绿体中Ca²⁺、ATP的结合	下调
	PK, pyk	编码丙酮酸激酶，叶绿体中糖酵解	上调
捕光叶绿素蛋白复合体	LHCP	编码叶绿素a/b结合蛋白-p，叶绿体类囊体膜上对光的吸收	下调
	LHCA1	编码叶绿素a/b结合蛋白-1，叶绿体类囊体膜上对光的吸收	下调
	LHCB4	编码叶绿素a/b结合蛋白-4，叶绿体类囊体膜上对光的吸收	下调
光合系统	PsaK	编码光系统Ⅰ(PSⅠ)-X亚基，光反应中心组份	下调
	PsbP	编码光系统Ⅱ(PSⅡ)放氧增强蛋白，反应中心组份	下调
	ATPF1D, atpH	编码F型H⁺运输ATP酶δ亚基，叶绿体类囊体膜质子运输	下调