生理指标和转录组测序分析脱落酸对葡萄果实色泽品质的影响

Physiological and transcriptomic analysis of effects of abscisic acid on grape fruit color quality

  • 摘要: 目的 基于生理指标和转录组测序分析脱落酸(ABA)对葡萄果实色泽品质的影响,为葡萄果实品质调控及葡萄产业高质量发展提供理论依据。方法 以园香妃、园金香、浪漫红颜和阳光玫瑰4个鲜食葡萄品种为试验材料,于葡萄果实转色初期,采用250 mg/L ABA对葡萄果穗进行浸果处理(T),不含ABA的溶液浸果处理为对照组,4个品种葡萄第1次采样标记为CK0,CK的第2、3次采样分别标记为CK1和CK2;ABA处理第2、3次采样分别标记为T1和T2。测定不同处理下不同品种葡萄的果实纵径、横径、粒重和可溶性固形物(TSS)、果皮花青苷、类胡萝卜素、叶绿素a、叶绿素b含量,并对园香妃葡萄果皮处理样本进行转录组测序分析,筛选差异表达基因(DEG),通过农杆菌瞬时转化巨峰葡萄果实检测关键基因表达情况。结果 与CK2相比,T2处理下,园香妃和园金香葡萄果实的纵径、横径和粒重均显著增加(P<0.05,下同),分别增加5.8%、8.3%、16.0%和8.9%、5.4%、16.5%;阳光玫瑰葡萄果实的横径和粒重显著增加,分别增加8.2%、21.7%。与CK2相比,T2处理园香妃和浪漫红颜葡萄果皮中总花青苷含量均极显著增加(P<0.001,下同),园香妃和浪漫红颜葡萄果皮5类花青苷组分中,均以T2处理含量最高,显著高于其他4个处理。与CK2相比,T2处理下园金香葡萄果皮类胡萝卜素、叶绿素a、叶绿素b含量均显著降低;浪漫红颜葡萄果皮叶绿素a含量显著降低;阳光玫瑰葡萄果皮类胡萝卜素和叶绿素a含量显著降低。转录组测序分析结果显示,在园香妃果皮样本中共检测到5379个DEGs。对DEGs进行GO功能注释分析,结果显示,T2 vs CK2组生物过程类别主要注释到温度刺激响应、机械刺激响应等GO功能条目。KEGG信号通路富集分析结果显示,T1 vs CK1组、T2 vs CK2组均在芪类、二芳基庚烷类及姜辣素生物合成、类黄酮生物合成等信号通路明显富集,与CK2相比,T2处理下园香妃葡萄果皮中苯丙氨酸解氨酶基因(PAL肉桂酸-4-羟化酶基因(C4H黄烷酮-3'羟化酶基因(F3'H)、黄酮醇合酶基因(FLS)、二氢黄酮醇-4-还原酶基因(DFR)与花青苷合成相关的DEGs表达水平整体明显上调。巨峰葡萄果实瞬时过表达试验结果显示,经R2R3-MYB转录因子基因(VvMYB24)过表达处理后,葡萄果皮花青苷含量显著增加,VvMYB24、查尔酮异构酶基因(VvCHI、VvDFR、葡萄白藜芦醇双加氧酶基因(VvLDOX)的相对表达量也极显著增加。结论 ABA通过促进花青苷合成基因VvPALVvC4H和转录因子基因VvMYBA1VvMYBA2VvMYBA24表达水平,抑制转录因子VvMYBC2-L1的表达水平促进葡萄果皮中花青苷的合成。

     

    Abstract: Objective This study aimed to investigate the effects of abscisic acid (ABA) treatment on color quality of grape fruits based on analysis of physiological indicators and transcriptome sequencing, so as to provide a theoretical basis for quality regulation and high-quality development of grape industry.Method Four table grape cultivars (Yuan Xiangfei, Yuan Jinxiang, Wagamichi and Shine Muscat) were used as materials. At initial veraison stage, the clusters were dipped in 250 mg/L ABA solution (T), and those clusters dipped in the solution without ABA were taken as control. The control group in the first sampling was labeled CK0; the control groups in the second and the third samplings were labeled CK1 and CK2; the ABA treatments in the second and third samplings were labeled T1 and T2. Fruit longitudinal dia-meter, transverse diameter, berry weight, total soluble solids (TSS), and contents of anthocyanins, carotenoids, chlorophyll a and chlorophyll b in berry skins of different grape cultivars were measured under different treatments. Transcriptome sequencing of grape skin samples of Yuan Xiangfei cultivar was performed to screen differentially expressed genes (DEGs). The expression of key genes was validated through Agrobacterium-mediated transient transformation in Kyoho grapes.Result Compared with CK2, T2 treatment significantly increased (P<0.05) the longitudinal diameter, transverse diameter, and berry weight of Yuan Xiangfei cultivar (by 5.8%, 8.3% and 16.0%) and Yuan Jinxiang (by 8.9%, 5.4% and 16.5%), while only transverse diameter and berry weight were significantly enhanced in Shine Muscat (by 8.2% and 21.7%). Compared with CK2, total anthocyanin content increased highly significantly (P<0.001,the same below) in grape skins of the Yuan Xiangfei and Wagamichi cultivars under T2 treatment. Among the five anthocyanin components in the grape skins of Yuan Xiangfei and Wagamichi, the highest accumulation was observed in T2, which was significantly higher that than in the other four groups. Compared with CK2, the contents of carotenoid, chlorophyll a and chlorophyll b were decreased significantly in Yuan Jinxiang, while chlorophyll a was reduced in Wagamichi, and both carotenoid and chlorophyll a were significantly decreased in Shine Muscat. Transcriptome analysis identified 5379 DEGs in the grape skin of Yuan Xiangfei. GO functional annotation analysis of DEGs revealed that in the group of T2 vs CK2, biological processes were mainly annotated to temperature and mechanical stimulus responses. KEGG signal pathway enrichment analysis showed that: in the groups of T1 vs CK1 and T2 vs CK2, significant enrichments were observed in stilbenoid, diarylheptanoid, gingerol and flavonoid biosynthesis; compared with CK2, in the Yuan Xiangfei cultivar, the anthocyanin-related DEGs such as the genes of phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase gene (C4H), flavanone-3'hydroxylase (F3'H), flavonol synthase (FLS), dihydroflavonol 4-reductase (DFR) gene exhibited obvious upregulated expressions. Transient overexpression of Kyoho grape fruits showed that, after overexpression treatment of MYB transcription factor gene (VvMYB24), the anthocyanin content in grape skin was significantly increased, and chalcone isomerase gene (VvCHI), VvDFR, grape resveratrol dioxygenase (VvLDOX) also showed a highly significantly increased related expression.Conclusion ABA promotes anthocyanin biosynthesis in grape skins by upregulating expression of anthocyanin biosynthesis-related genes (VvPALVvC4H) and transcription factor genes (VvMYBA1VvMYBA2VvMYBA24) while inhibiting the expression of the transcription factor (VvMYBC2-L1).

     

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