异源授粉山核桃果皮光合能力差异的转录组分析

doi:10.11707/j.1001-7488.20190115

摘要/Abstract

摘要： [目的]探讨采用薄壳山核桃花粉授粉增强山核桃果皮光合能力的分子机理，为花粉直感的深入研究奠定基础。[方法]以山核桃花粉（记为hp）和薄壳山核桃花粉（记为pp）授粉的山核桃果皮为研究材料，在山核桃果实快速膨大期（授粉后65天，65DAP），对不同花粉授粉后山核桃果皮进行转录组测序，通过对叶绿素合成、光反应、碳同化等通路的基因富集分析，结合叶绿素含量、光合速率及光合关键酶活性的变化，筛选出增强山核桃果皮光合能力的相关基因。[结果]65DAP时，pp授粉的山核桃果皮中叶绿素含量、光合速率、1，5-二磷酸核酮糖羧化酶（Rubisco）活性、磷酸烯醇式丙酮酸羧化酶（PEPC）活性分别是hp授粉的1.31倍（P=0.047）、1.12倍（P=0.000 43）、1.65倍（P=0.036）和1.23倍（P=0.001 3）。转录组测序共产生32 908条scaffolds，并从1 894个差异基因（P<0.05，foldchange>1.5倍）中筛选出66个与山核桃果皮光合相关的基因，主要富集在叶绿素合成通路及光合固碳途径中，其中叶绿素合成途径中镁螯合酶编码基因（CHLH）、镁-原卟啉单甲酯环化酶编码基因（CRD）和叶绿素b还原酶编码基因（NYC）表达量明显上调，脱酶叶绿酸编码基因（PAO）表达量则显著下降；光合碳同化途径中有16个基因以及Rubisco活化酶编码基因（RCA）和碳酸酐酶编码基因（CA）表达量均明显上调；与光保护相关的42个基因表达量也明显上调。[结论]在山核桃果实快速膨大期，薄壳山核桃花粉授粉的山核桃果皮中的镁螯合酶编码基因（CHLH）、镁-原卟啉单甲酯环化酶编码基因（CRD）、叶绿素b还原酶编码基因（NYC）、捕光蛋白编码基因（LHCⅡ）、光修复相关蛋白编码基因（PSAN、PSAB27、STN7）和38个热激蛋白编码基因（HSP）、Rubisco活化酶编码基因（RCA）以及碳酸酐酶编码基因（CA）均显著上调，表明薄壳山核桃授粉的山核桃果皮光合能力的增强与其叶绿素合成、光能捕获和碳固定等光合作用相关通路的基因表达上调有关。

关键词: 山核桃, 薄壳山核桃, 花粉直感, 光合能力, 基因差异表达

Abstract: [Objective] To elucidate the molecular mechanism for enhancing photosynthetic capacity of the exocarp of Carya carthayensis fruits pollinated by Carya illinoensis in order to provide a theoretic basis for further research on metaxenia.[Method] Transcriptome sequencing was performed for exocarp of C. cathayensis respectively pollinated by C. cathayensis (marked as hp) and C. illinoensis (marked as pp) with samples collected 65 days after pollination (65DAP) when the fruit rapidly expanding in size. Gene enrichment analysis was conducted for pathways of chlorophyll synthesis, photo-response and carbon assimilation in combination with variation in chlorophyll content, photosynthetic rate and activity of key enzymes involved in the photosynthesis.[Result] Compared to the fruit pollinated with hp, the chlorophyll content, photosynthetic rate, Rubisco activity, and phosphoenolpyruvate carboxylase (PEPC) activity of exocarp in the fruit pollinated with pp were 1.31 (P=0.047), 1.12 (P=0.000 43), 1.65 (P=0.036), and 1.23 (P=0.001 3) folds, respectively. A total of 32 908 scaffolds were generated by transcriptome sequencing, and 66 photosynthesis-related genes were screened from 1 894 differentially expressed genes (DEGs) (P <0.05,fold change>1.5), which were mainly involved in pathways of chlorophyll synthesis and photosynthetic carbon fixation. The expression of magnesium-chelatase subunit (CHLH), agnesium-protoporphyrin IX monomethyl ester cyclase (CRD), and chlorophyll(ide) b reductase(NYC) of the fruit pollinated with pp was significantly up-regulated while that of pheophorbide a oxygenase (PAO) was significantly down-regulated in the chlorophyll biosynthesis pathway, and the expression of 16 light harvesting pigment protein complexes (LHCⅡ)-coding genes, ribulose bisphosphate carboxylase/oxygenase activase (RCA), alpha carbonic anhydrase (CA) in the photosynthetic carbon fixation pathway were significantly up-regulated. 42 genes involved in photoprotection of exocarp are also showed significantly high expression in the fruit pollinated with pp than those in the fruit pollinated with hp.[Conclusion] In the rapid growth period of Carya carthayensis fruits, the expression of magnesium-chelatase subunit (CHLH), agnesium-protoporphyrin IX monomethyl ester cyclase (CRD), chlorophyll(ide) b reductase (NYC), light harvesting pigment protein complexes-coding genes (LHCⅡ), light repair protein-coding genes (PSAN, PSAB27, STN7), 38 heat shock proteins (HSP), ribulose bisphosphate carboxylase/oxygenase activase (RCA) and alpha carbonic anhydrase (CA) was significantly up regulated for the pp cross combination. It indicates that the significantly up-regulated expression of genes involved in chlorophyll synthesis, light capture and carbon assimilation might be related to the significantly higher photosynthetic rate in the exocarp of pp cross combination.

Key words: Carya cathayensis, Carya illinoensis, metaxenia, photosynthetic capacity, differential expression of gene

中图分类号:

S718.46

黄仁, 张韵, 张启香, 王正加, 夏国华, 黄坚钦, 胡渊渊. 异源授粉山核桃果皮光合能力差异的转录组分析[J]. 林业科学, 2019, 55(1): 128-137.

Huang Ren, Zhang Yun, Zhang Qixiang, Wang Zhengjia, Xia Guohua, Huang Jianqin, Hu Yuanyuan. Transcriptome Analysis of Photosynthetic Capacity of Exocarp of Heterogeneously Pollinated Carya cathayensis[J]. Scientia Silvae Sinicae, 2019, 55(1): 128-137.

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