Transcriptomics and metabolomics analyses provide insights into postharvest ripening and senescence of tomato fruit under low temperature

Tomato is one of the most important vegetable crops in the world and is a model plant used to study the ripening of climacteric fleshy fruit.During the ripening process of tomato fruit,flavor and aroma metabolites,color,texture and plant hormones undergo significant changes.However,low temperatures delayed the ripening process of tomato fruit,inhibiting flavor compounds and ethylene production.Metabolomics and transcriptomics analyses of tomato fruit stored under low temperature(LT,5°C)and room temperature(RT,25°C)were carried out to investigate the effects of storage temperature on the physiological changes in tomato fruit after harvest.The results of transcriptomics changes revealed that the differentially expressed genes(DEGs)involved in tomato fruit ripening,including several kinds of transcription factors(TFs)(TCP,WRKY,MYB and bZIP),enzymes involved in cell wall metabolism[beta-galactosidase(β-GAL),pectinesterase(PE)and pectate lyase(PL),cellulose and cellulose synthase(CESA)],enzymes associated with fruit flavor and aroma[acetyltransferase(AT),malic enzyme(ME),lipoxygenase(LOX),aldehyde dehydrogenase(ALDH),alcohol dehydrogenase(ADH)and hexokinase(HK)],genes associated with heat stress protein 70 and genes involved in the production of plant hormones such as Ethylene responsive factor 1(ERF1),Auxin/indoleacetic acids protein(AUX/IAA),gibberellin regulated protein.Based on the above results,we constructed a regulatory network model of the effects of different temperatures during the fruit ripening process.According to the analysis of the metabolomics results,it was found that the contents of many metabolites in tomato fruit were greatly affected by storage temperature,including,organic acids(L-tartaric acid,a-hydroxyisobutyric acid and 4-acetamidobutyric acid),sugars(melezitose,beta-Dlactose,D-sedoheptulose 7-phosphate,2-deoxyribose 1-phosphate and raffinose)and phenols(coniferin,curcumin and feruloylputrescine).This study revealed the effects of storage temperature on postharvest tomato fruit and provided a basis for further understanding of the molecular biology and biochemistry of fruit ripening.

ERF4 affects fruit ripening by acting as a JAZ interactor between ethylene and jasmonic acid hormone signaling pathways

The regulation of apple(Malus domestica)fruit texture during ripening is complex and a fundamental determinant of its commercial quality.In climacteric fruit,ripening-related processes are regulated by ethylene(ET),and jasmonate(JA)is also involved in the ethylene biosynthesis pathway,mainly through the transcription factor MYC2.However,the molecular genetic mechanism for fruit ripening processes between the JA and ET signaling pathways still needs to be elucidated.In order to explore how JA regulates apple fruit ripening through ERF4,we used’Gala’and’Ralls Janet’fruit at different developmental stages as experimental materials to determine the fruit firmness and related gene expression analysis.Meanwhile,we carried out different hormone treatments on’Gala’fruit at ripening stage.Here,we show that ERF4 is a core JA signaling hub protein JASMONATE ZIM-DOMAIN(JAZ)interactor that affects ethylene signaling pathways.During fruit development,ERF4 represses the expression of ACS1 and ACO1 by interacting with JAZ,as well as with the JA-activated transcription factor MYC2.Ripening is promoted in JAZ-suppressed apples.Thus,ERF4 acts as a molecular link between ethylene and JA hormone signals,and the natural variation of the ERF4Ethylene-responsive binding factor-associated amphiphilic repression(EAR)motif decreases repression of ethylene biosynthesis genes.

Transcriptional profiles underlying the effects of salicylic acid on fruit ripening and senescence in pear(Pyrus pyrifolia Nakai)

Salicylic acid(SA) plays a pivotal role in delaying fruit ripening and senescence. However, little is known about its underlying mechanism of action. In this study, RNA sequencing was conducted to analyze and compare the transcriptome profiles of SA-treated and control pear fruits. We found a total of 159 and 419 genes differentially expressed between the SA-treated and control pear fruits after 12 and 24 h of treatment, respectively. Among these differentially expressed genes(DEGs), 125 genes were continuously differentially expressed at both treatment times, and they were identified as candidate genes that might be associated with SA-regulated fruit ripening and senescence. Bioinformatics analysis results showed that 125 DEGs were mainly associated with plant hormone biosynthesis and metabolism, cell wall metabolism and modification, antioxidant systems, and senescence-associated transcription factors. Additionally, the expression of several candidate DEGs in ripening and senescent pear fruits after SA treatments were further validated by quantitative real-time PCR(qRT-PCR). This study provides valuable information and enhances the understanding of the comprehensive mechanisms of SA-meditated pear fruit ripening and senescence.

PaPYL9 is involved in the regulation of apricot fruit ripening through ABA signaling pathway

Abscisic acid(ABA)is a major regulator of non-climacteric fruit ripening;however,the role of ABA in the ripening of climacteric fruit is not clear.Here,as a typical climacteric fruit,apricots were used to investigate the role of ABA in fruit ripening.Based on weighted gene coexpression network analysis(WGCNA)of our previous transcriptome data,we treated‘Danxing’fruit with exogenous ABA and obtained ABA receptor genes,genes related to ABA biosynthesis and signal transduction,and analyzed the response of these candidate genes to exogenous ABA during fruit ripening.Subsequently,the full length of candidate PYLs genes were cloned,and their putative function were analyzed by phylogenetic analysis and protein structure domain analysis.And then the function of one candidate gene PaPYL9 was verified by using transgenic tomato.Furthermore,the response genes in transgenic tomato were screened by transcriptome sequencing,and ultimately the related regulatory network was proposed.The results showed that the injection of exogenous 1.89 mmol·L^(-1) ABA remarkably promoted fruit coloration,and increased the color index for red grapes(CIRG)and the total soluble solids(TSS)content,but significantly decreased the firmness and titratable acid(TA)content(p<0.01).Nordihydroguaiaretic acid(NDGA),the inhibitor of ABA,appeared to have the converse role in TA,TSS,CIRG and firmness,during the ripening process.One NCED(9-cis-epoxycarotenoiddioxygenase)and five ABA receptor genes related to signal transduction were mined from the transcriptome data of apricot fruit through WGCNA.Compared with the control,the expression levels of NCED1,PYL9(PYR/PYL/RCAR),SnRK2(SUCROSE NON-FERMENTING1(SNF1)-RELATED PROTEIN KINASE 2S),and ABF2(ABRE-binding bZIP transcription)were induced dramatically by ABA treatment(p<0.01),while NDGA treatment significantly inhibited their expression.Based on gene expression and protein domain analysis,we inferred that PaPYL9 is putatively involved in apricot fruit ripening.Overexpression of PaPYL9 in Micro-TOM tomatoes resulted in the promotion of early ripening.Simultaneously,the expression levels of genes related ethylene biosynthesis,chlorophyll degradation,fruit softening,flavor formation,pigment synthesis,and metabolism were all significantly induced in overexpression of PaPYL9 tomatoes.This indicates the central role of ABA in climacteric fruit ripening.A regulatory network was tentatively proposed,laying the foundation to unveil the molecular mechanism of the regulatory role of PaPYL9 in fruit ripening.

Changes of Reactive Oxygen Species and Related Enzymes in Mitochondria Respiratory Metabolism During the Ripening of Peach Fruit

The fruits of peach cultivar Yuhua 3 were used as materials to investigate the changes of active oxygen and related enzymes in mitochondria respiratory metabolism during ripening of peach fruit, involving their influence on the proceeding of peach fruit senescence. The results showed that the large decrease in firmness occurred between maturity II and IV. The decrease in firmness coincided with an increase in respiratory intensity. Obvious peaks of respiratory intensity lagging to the rapid change of fruit firmness could be shown during peach ripening. Reactive oxygen species （ROS） had a cumulative process and positively correlated with respiratory intensity. During peach ripening, the content of Ca^2＋ increased, the activities of succinic dehydrogenase （SDH）, cytochrome C oxidase （CCO）, H＋-ATPase, and Ca^2＋-ATPase decreased varying in different degree at the later step of ripening. These suggested a close relationship existed between ROS metabolism and mitochondrial respiration, namely, both ROS metabolism and mitochondrial respiration probably played important roles in ripening and senescing of peach fruit.

Changes of Activities in NAD Kinase and NADP PhosphataseDuring Ripening and Senescence of Tomatoand Strawberry Fruits

Activities of NAD kinase(NADK)and NADP phosphatase and relationship between the two enzymes and temperature, respiration, ethylene production and trifluoperazine(TFP) were studied during ripening and senescence of strawberry and tomato fruits after harvest at 4℃ and 20℃. The activity of NAD kinase in strawberry decreased slowly during first four days, then increased gradually. The NADP phosphatase activity increased at the second day, decreased the next day,then increased again. In tomato fruit, the activities of NAD kinase and NADP phosphatase increased at the second day, decreased with the ripening and senescence of the fruit. The change trend of NAD kinase and respiration in the two fruits were similar, the same were NADP phosphatase and ethylene production. TFP enhanced the activity of NAD kinase and had little effect on NADP phosphatase. Low temperature(4℃) activated the NAD kinase and reduced the activity of NADP phosphatase. These results indicated that the NAD kinase and NADP phosphatase were related to the ripening and senescence of strawberry and tomato fruits. The activation of NAD kinase probably postponed the ripening and senescence of the fruits.

Characteristics of fruit ripening in tomato mutant epi

The characteristics of fruit ripening and expression of ripening-related genes were investigated in epi, an ethylene overproduction mutant of tomato (Lycopersicon esculentum Mill.). The epi produces apparently more ethylene than its wild type VFN8 at every stage of vegetative and fruit growth and ripening; compared to VFN8, the epi fruit showed higher CO2 evolution, faster descending of chlorophyll, slightly quicker increase of carotenoid and lycopene, and faster reduction in pericarp firmness during maturation and ripening; and the mRNAs of three ripening-related genes including E8, pTOM5 and pTOM6 were at higher levels in epi. The ripening-related characteristics changing of the fruit are consistent with the increase of ethylene production and ripening-related genes expression. These results suggest that epi mutation possibly did not affect the ethylene perception and signaling during fruit ripening, and that the modified characteristics of fruit ripening possibly resulted from the ethylene over- production and increased expression of ripening-related genes.

Elucidating the role of SlXTH5 in tomato fruit softening

Fruit softening in tomato(Solanum lycopersicum)is closely associated with cell wall disassembly,which is brought about through the action of a range of cell wall structure-related enzymes and other proteins such as expansins.Xyloglucan endotransglucosylase/hydrolase(XTH)(EC 2.4.1.207 and/or EC 3.2.1.151)has been proposed to be key player involved in xyloglucan metabolism.SlXTH5 showed the highest expression level among all SlXTHs during tomato ripening.In this study,the role of SlXTH5 involved in tomato softening was investigated in CRISPR-based knockout mutants of SlXTH5.Loss-of-function of SlXTH5 in transgenic tomato lines resulted in slightly firmer fruit pericarp,but significantly decreased their color index compared with azygous wild type(WT)control fruits.Increased paste viscosity was detected in CRISPR mutants,indicating that the activity of SlXTH5 is responsible for maintaining cell wall structural integrity.Immunocytochemistry studies were performed using the monoclonal antibody probe LM25 to examine the localization and distribution of xyloglucan in the pericarp cells of the CRISPR mutant fruits.The data indicated more xyloglucan was retained in the pericarp of CRISPR mutant fruit than in WT control fruit.This study revealed the link between SlXTH5 and xyloglucan metabolism and indicated the potential of manipulating SlXTH5 to regulate fruit softening.

Over-expression of GhDWF4 gene improved tomato fruit quality and accelerated fruit ripening

Brassinosteroids（BRs）, a class of steroidal phytohormones are essential for many biological processes in plant. However, little is known about their roles in fruit development. Tomato is a highly valuable vegetable and has been adopted as the model species for studying fruit growth, development, and ripening. To understand the role of endogenous BRs in the development of tomato fruit, the expression patterns of three homologues of DWF4 gene were investigated and the transgenic tomato plants were generated in which the Gh DWF4 gene from upland cotton（Gossypium hirsutum L.） was ectopically expressed. The contents of main quality components were analyzed in fruits of transgenic tomato line and non-transgenic line（control plant, CP） when the fruit was mature. Sl CYP90B3 that possesses high homology with Gh DWF4 preferentially expressed in mature fruit. Significantly higher contents of soluble sugar, soluble proteins, and vitamin C were obtained in fruit of transgenic tomato lines compared with those in the CP. Furthermore, overexpressing Gh DWF4 promoted fruit growth and ripening. The weight per fruit was increased by about 23% in transgenic lines. In addition, overexpressing Gh DWF4 promoted the germination of transgenic tomato seeds and hypocotyl elongation of seedlings. These results indicated that overexpressing Gh DWF4 gene in tomato could increase the contents of many nutrients in fruit and accelerate fruit ripening. It is suggested that increased endogenous BRs in fruit affect the growth and development of tomato fruit and therefore improved the nutrient quality of tomato.

The Influence of Co-Suppressing Tomato 1-Aminocyclopropane-1-Carboxylic Acid OxidaseⅠon the Expression of Fruit Ripening-Related and Pathogenesis-Related Protein Genes

The purpose of this study is to explore the influence of co-suppressing tomato ACC oxidase Ⅰ on the expression of fruit ripening-related and pathogenesis-related protein genes, and on the biosynthesis of endogenous ethylene and storage ability of fruits. Specific fragments of several fruit ripening-related and pathogenesis-related protein genes from tomato （Lycopersicon esculentum） were cloned, such as the l-aminocyclopropane-1-carboxylic acid oxidase 1 gene （LeAC01）, 1- aminocyclopropane-l-carboxylic acid oxidase 3 gene （LeAC03）, EIN3-binding F-box 1 gene （LeEBF1）, pathogenesis-related protein 1 gene （LePR1）, pathogenesis-related protein 5 gene （LePR5）, and pathogenesis-related protein osmotin precursor gene （LeNP24） by PCR or RT-PCR. Then these specific DNA fragments were used as probes to hybridize with the total RNAs extracted from the wild type tomato Ailsa Craig （AC＋＋） and the LeAC01 co-suppression tomatoes （V1187 and T4B）, respectively. At the same time, ethylene production measurement and storage experiment of tomato fruits were carded out. The hybridization results indicated that the expression of fruit ripening-related genes such as LeACO3 and LeEBF1, and pathogenesis-related protein genes such as LePR1, LePR5, and LeNP24, were reduced sharply, and the ethylene production in the fruits, wounded leaves decreased and the storage time of ripening fruits was prolonged, when the expression of LeACO1 gene in the transgenic tomato was suppressed. In the co-suppression tomatoes, the expression of fruit ripening-related and pathogenesis-related protein genes were restrained at different degrees, the biosynthesis of endogenous ethylene decreased and the storage ability of tomato fruits increased.

Comparative transcriptome analysis of the climacteric of apple fruit uncovers the involvement of transcription factors affecting ethylene biosynthesis

Apple(Malus domestica)fruit generally undergoes a climacteric.During its ripening process,there is a peak in ethylene release and its firmness simultaneously decreases.Although more in-depth research into the mechanism of climacteric-type fruit ripening is being carried out,some aspects remain unclear.In this study,we compared the transcriptomes of 0-Pre and 15-Post(pre-and post-climacteric fruit),and 15-Post and 15-MCP[fruit treated with 1-MCP(1-methylcyclopropene)].Various transcription factors,such as MADS-box,ERF,NAC,Dof and SHF were identified among the DEGs(differential gene expressions).Furthermore,these transcription factors were selected for further validation analysis by qRT-PCR.Moreover,yeast one hybrid(Y1H),β-glucuronidase(GUS)transactivation assay and dual-luciferase reporter assay showed that MdAGL30,MdAGL104,MdERF008,MdNAC71,MdDof1.2,MdHSFB2a and MdHSFB3 bound to MdACS1 promoter and directly regulated its transcription,thereby regulating ethylene biosynthesis in apple fruit.Our results provide useful information and new insights for research on apple fruit ripening.

优质大果中熟白肉枇杷新品种中白的选育

中白是从早钟6号×贵妃杂交群体中选育的优质大果中熟白肉枇杷新品种。该品种树势中庸偏强,树冠圆头形,中心干较明显,枝梢较强壮。平均单果质量60.1 g,大小整齐;果实倒卵圆形,果基钝圆少数尖峭,果顶钝圆微凹;果皮橙黄色,厚度中等偏薄,锈斑少,果点密度中等,果点中等大;萼片平展,萼孔闭合;果肉黄白色,平均厚度10.9 mm,肉质细嫩、化渣、汁多;平均可溶性固形物含量(w)13.4%,味鲜、风味浓郁,品质优;平均可食率71.2%,平均种子4.6粒·果^(-1);在福建福州地区,果实4月中下旬成熟。高接换种后第2年可少量开花,第3年枝梢抽穗率50%左右,第4年株产22.8 kg,折合666.7 m^(2)产709.7 kg,早结、丰产、稳产。中白是一个优质、大果、丰产稳产、抗性较强的中熟白肉枇杷新品种,适宜在福建、四川、重庆、云南等枇杷产区应用。

猕猴桃果实淀粉代谢研究进展

猕猴桃为国际重要水果种类,已成为我国精准扶贫、乡村振兴的“金果果”。淀粉作为植物光合作用固定碳形成的主要碳水化合物,在植物的整个生长发育过程中具有重要作用。猕猴桃果实属于淀粉积累型水果,在临近商业采收时淀粉积累达到峰值,然后随着果实软化成熟淀粉降解为糖,果实甜度增高,风味品质形成。同时,猕猴桃属于呼吸跃变型果实,具有生理后熟属性,采后易软化,不耐贮藏。淀粉作为细胞内容物对维持细胞膨压,支持果实硬度起着重要作用。随着猕猴桃基因组的测序完成,猕猴桃果实淀粉代谢分子研究取得新的进展,尤其是淀粉降解方面,但是目前对猕猴桃果实淀粉代谢进展的整理与归纳还鲜有报道。因此,从猕猴桃果实淀粉的理化性质、植物淀粉代谢途径以及猕猴桃果实淀粉代谢分子机制三个方面展开,并结合淀粉与猕猴桃果实风味品质、成熟软化的关系,对猕猴桃果实淀粉研究现状与进展进行综述,为以后创制优质高淀粉耐贮藏猕猴桃新材料或选育新品种,以及建立即食供应的快速后熟技术体系提供理论支撑。

桃基因 PpNAC 的鉴定及其在不同发育时期的表达分析

【目的】鉴定影响桃果实成熟的PpNAC基因,并分析其在桃果实不同发育时期的表达量变化,为解析桃果实成熟的分子机制奠定基础。【方法】通过对桃不同组织部位的转录组分析,从115个PpNAC基因家族成员中筛选在果实中特异表达的PpNAC基因,进一步进行生物信息学分析、蛋白互作分析和表达分析,筛选可能影响桃果实成熟的PpNAC基因。【结果】筛选到16个在桃果实中高表达的PpNAC基因,其在桃的8条染色体上都有分布,蛋白相对分子质量和等电点差异较大。基因编码区均含有NAM保守结构域和motif1~6,含有3~8个外显子,2~7个内含子,启动子中脱落酸响应元件和茉莉酸响应元件相对较多。此外,通过与已报道成熟相关NAC蛋白PpNAC1的互作筛选,得到4个与PpNAC1互作的PpNAC蛋白。进一步的表达分析表明,这4个PpNAC基因在果实成熟起始期和成熟后期表达量达到最高。【结论】PpNAC基因可能参与调控桃果实成熟。

香蕉AP2/ERFs超家族的重新鉴定及在果实采后成熟过程中的差异表达特性

【目的】在全基因组水平上重新鉴定香蕉A基因组中的AP2/ERF家族成员,研究其在香蕉果实采后成熟过程中的差异表达特性,明确可能参与香蕉果实成熟调控的关键基因。【方法】对香蕉A基因组中AP2/ERF家族成员进行系统进化、结构特征、蛋白质特性、保守结构域分析和两大类主栽品种巴西蕉(AAA)和粉蕉(ABB)果实采后成熟不同阶段的转录组分析。【结果】发现AP2/ERFs家族共有317个家族成员,分为AP2(49个)、ERF(253)和RAV(15)三个亚家族,他们不均匀地分布在染色体上。根据保守结构域和基因结构特征,ERF又分为a、b、c、d、e、f、h、i、j和k共10个亚类。转录组分析结果表明,在巴西蕉果实采后成熟过程中差异表达的AP2/ERFs家族成员有77个,其中高水平表达的有MaERF15、36、42和AP2-28。在粉蕉果实采后成熟过程中差异表达的AP2/ERFs家族成员有74个,其中高水平表达的有MaERF42和AP2-28。同时在巴西蕉和粉蕉果实成熟过程中差异表达的基因有57个,其中高水平表达的基因有MaERF15、42和AP2-28,只在巴西蕉果实中特异表达的有20个,只在粉蕉中特异表达的有17个。【结论】重新鉴定了香蕉AP2/ERFs超家族成员及其在果实后熟过程中的差异表达特性,为系统深入解析香蕉AP2/ERF基因功能奠定了基础,对为调控香蕉果实成熟提供靶标基因具有一定的理论意义。

桑椹成熟相关脱落酸合成和信号转导关键基因的筛选与鉴定

脱落酸(ABA)是调控果实成熟的重要植物激素,在跃变型和非跃变型果实成熟中均发挥关键的调控作用。基于桑椹转录组数据筛选ABA合成和信号转导相关基因,结合基因表达和桑椹成熟相关生理指标鉴定重要基因,并分析基因功能。结果从筛选到的38个基因中,鉴定到9个在果实成熟中与ABA生物合成和信号转导相关的重要基因,其中4个基因LOC21404497(NCED)、LOC21384903(PP2C)、LOC21396283(SnRK2)、LOC21410137(ABF)在桑椹成熟和品质形成的关键时期(T4期)具有较高的表达水平,并且这4个基因的表达水平与桑椹硬度、酸度和可溶性固形物含量等指标具有高度相关性,表明这4个基因可能是桑椹成熟相关的ABA合成和信号转导的关键基因,在桑椹成熟调控中发挥重要作用。上述研究结果为深入探究桑椹成熟的分子调控机制提供了依据。

黑莓1-氨基环丙烷-1-羧酸氧化酶基因RuACO 1a和RuACO 1b的克隆及功能鉴定

基于前期黑莓(Rubus spp.)品种‘Navaho’果实转录组测序结果,通过反转录PCR克隆获得2个1-氨基环丙烷-1-羧酸氧化酶(ACO)基因,命名为RuACO 1a和RuACO 1b。结果表明:RuACO 1a和RuACO 1b的开放阅读框(ORF)长度分别为939和918 bp,分别含有4和3个外显子。系统进化分析结果显示RuACO1a和RuACO1b与月季(Rosa chinensis Jacq.)、蕨麻〔Argentina anserina(Linn.)Rydb.〕和野草莓(Fragaria vesca Linn.)ACO1蛋白的亲缘关系均较近,且分别属于蔷薇科(Rosaceae)植物中2类ACO1蛋白。RuACO 1a在果实着色后28 d响应乙烯利诱导,其相对表达量急剧升高,而RuACO 1b在果实着色后21 d响应乙烯利诱导,早于RuACO 1a。脱落酸处理后,RuACO 1a和RuACO 1b的相对表达量在果实着色后14和28 d较高。RuACO 1a和RuACO 1b具有组织表达特异性,RuACO 1a在幼果、花蕾和花中的相对表达量较高,RuACO 1b在幼果和幼根中的相对表达量较高。与野生型相比,RuACO 1a和RuACO 1b过量表达转基因拟南芥〔Arabidopsis thaliana(Linn.)Heynh.〕植株均表现为叶片中叶绿素相对含量和氮含量升高、开花和角果成熟提前、ACO含量升高。综上所述,黑莓RuACO 1a和RuACO 1b基因均促进拟南芥角果提前成熟。

植物甲基转移酶在果实成熟和品质形成中的作用研究进展

果实的成熟过程是色泽、香气、质地、营养等品质形成的关键阶段,受到多因子多层次的精细调控。甲基转移酶是一类在植物中普遍存在且功能广泛的重要酶类,部分甲基转移酶参与了果实成熟相关基因的表达调控,通过表观遗传修饰影响果实品质的形成,部分甲基转移酶广泛参与了多种生理生化过程,例如催化重要代谢物质的甲基化,改变其生理活性,从而影响植物的生长发育和果实成熟过程。为了进一步探究甲基转移酶对果实成熟和品质形成的调控作用,对甲基转移酶的分类方式进行了总结,详细综述了甲基转移酶在果实成熟和品质形成中的作用,并对相关甲基转移酶的研究方向加以展望,以期为进一步研究果实成熟及其品质形成机制提供参考。

设施栽培甘平杂柑果实枯水和浮皮调查与分析

为明确“甘平”杂柑果实枯水和浮皮症状与果实品质性状关系,进而为生产上的果实管理提供指导,以保温大棚内正常管理的5年生“甘平”树为研究对象,于2月下旬采收期进行整株采摘,对其浮皮和枯水发生情况进行调查,并逐一测定单个果实的品质性状,研究果实浮皮与不浮皮之间、枯水与不枯水之间品质性状的差异。结果表明,保温大棚内常规管理的“甘平”枯水发生率为59.8%,浮皮发生率为80.3%,两种症状的发生没有明显的相关性。不枯水果实的纵径、横径和单果质量显著高于枯水果实,表现为大果不容易枯水。浮皮果实横径、纵径、单果质量、果肉质量、可食率、果汁率、可溶性固形物含量和固酸比等显著高于不浮皮果实,果形指数显著小于不浮皮果实,表现为果实越大、果形越扁、品质越好的果实越容易出现浮皮。“甘平”枯水发生在果实膨大阶段。浮皮发生初期的果实品质较好,是完熟的表现。生产上,应注重合理负载,培养大果,并实行完熟栽培,以减少枯水的发生和提高果实品质。

葡萄CYP707A基因家族的鉴定及对果实成熟的功能验证

【目的】CYP707A是细胞色素P450家族的亚家族成员,所编码的ABA8'-羟化酶是ABA分解代谢的关键酶,对植物生长发育起着重要作用。鉴定葡萄CYP707A基因家族,可为进一步研究葡萄果实成熟机理和生物育种技术提供理论依据。【方法】利用生物学信息方法分析葡萄中CYP707A家族基因的基因结构、蛋白基序、染色体定位、共线性、系统进化关系及启动子区顺式作用元件等,并利用RT-q PCR分析VvCYP707A基因在葡萄不同品种(早熟和中熟)的时空表达模式、组织表达模式以及对外源ABA处理的响应模式,并利用果实瞬时表达技术对Vitvi07g01751进行功能验证。【结果】葡萄基因组鉴定出5个VvCYP707As基因,分布于5条染色体,其中Vitvi02g01269、Vitvi07g01751和Vitvi18g00792为片段复制基因。CYP707A基因家族高度保守,其中,基因结构和蛋白基序相似,亚细胞定位均位于内质网;VvCYP707A启动子区域富含与生长发育和激素响应相关的顺式作用元件,均含有ABA激素响应元件;通过RT-qPCR检测基因的表达模式发现,VvCYP707A基因具有时空特异性和组织特异性,主要在果实的幼果期与膨大期和茎、叶组织中高表达,其中Vitvi07g01751基因在早熟品种‘甬早红’膨大期果实中特异性高表达,且在两个品种间的表达模式一致,随着果实的成熟其表达先升高后降低。外源ABA处理下VvCYP707A分为两种表达模式,在浆果肉中外源ABA可诱导5个VvCYP707A基因高表达,在浆果皮中会显著抑制VvCYP707A基因的表达;在葡萄果实中瞬时表达Vitvi07g01751,葡萄的糖度积累缓慢,显著增加编码ABA的受体VvPYL4基因表达量,推测了Vitvi07g01751介导ABA调控葡萄果实成熟的机制。【结论】研究结果揭示了葡萄Vitvi07g01751在果实成熟中起着重要作用。