‘仓方早生’桃及其早熟芽变不同发育时期果实的转录组分析

【目的】通过对桃品种‘仓方早生’及其早熟芽变不同发育时期的果实进行转录组分析，挖掘参与调控桃果实成熟的关键因子，为深入研究桃果实成熟调控机理提供理论依据。【方法】以桃品种‘仓方早生’及其早熟芽变为试材，每个品种分别选择长势一致的样品树5株，分别于花后30 d（对应‘仓方早生’c1、早熟芽变y1）、45 d（对应c2、y2）、59 d（对应c3、y3）、71 d（对应c4、y4）及89 d（对应c5）对不同发育时期的桃去皮果肉进行取样和转录组测序，并利用实时荧光定量PCR(qRT-PCR)对筛选的差异表达基因进行定量验证；利用GO和KEGG对‘仓方早生’及其早熟芽变的差异表达基因进行分析；基于差异表达基因构建加权基因共表达网络分析（weighted gene co-expression network analysis,WGCNA），从中鉴定出与果实成熟密切相关的枢纽模块和枢纽基因。【结果】将处于果实相同发育时期的转录组数据进行比较，得到y1与c1、y2与c2、y3与c4和y4与c5四组对比数据，共筛选出差异表达基因4 395个，其中上调表达基因2 212个，下调表达基因2 183个。其...     

1-MCP和SO2保鲜剂处理阳光玫瑰葡萄的转录组学分析

【目的】分析2种保鲜剂处理阳光玫瑰葡萄贮藏阶段基因差异表达情况，为从分子生物学角度研究保鲜剂处理对葡萄果实贮藏品质影响的调控机制提供理论参考。【方法】以阳光玫瑰葡萄为试材，采用1-甲基环丙烯（1-MCP）和二氧化硫（SO₂）保鲜剂分别对果实进行冰温贮藏（-1~1℃），并以不使用保鲜剂的果实为对照，对贮藏1、5、9、13和17周的2种保鲜剂处理及对照的果实分别取样开展转录组学分析，通过实时荧光定量PCR结果证实转录组测序结果的可靠性。【结果】从对照和2种保鲜剂处理的转录组测序结果中共获得371.64 Gb的原始数据，各样品Clean data均达6.03 Gb，GC含量为46.28%~47.53%，Q30≥93.50%，与葡萄参考基因组的匹配率为75.75%~90.23%。1-MCP处理与对照的差异表达基因数在贮藏后13周达最高值，而SO₂处理与对照及SO₂处理与1-MCP处理的差异表达基因数在贮藏后5周达最高值。贮藏1周和13周时，1-MCP处理与对照之间差异表达基因最多，分别为965和2881个；贮藏5周和9周时，SO₂处理与对照之间差异表达基因最多，分别为3698和1628个；贮藏17周时，处理和对照两两比较获得的差异表达基因相差不大。贮藏1~5周果实内部发生细胞组分、分子功能和生物过程等方面的变化较贮藏中后期更为剧烈，且在MYB、AP2/ERF-ERF、NAC、GARP-G2-like、HB-HD-ZIP和WRKY转录因子的调控下，单萜合成相关结构基因表达量在贮藏5周后迅速降低。基于实时荧光定量PCR的所有样本中苯丙烷—类黄酮、类胡萝卜素和单萜合成代谢路径相关基因表达水平检测结果与转录组测序分析结果基本一致。【结论】 1-MCP与SO₂保鲜剂对阳光玫瑰葡萄贮藏产生不同影响，前者在贮藏过程中抑制果实成熟和衰老作用释放较后者更为平缓，且通过阻断乙烯与受体结合并抑制ETR、EIN3和ERF1/2这3个乙烯信号通路关键基因表达发挥延缓衰老作用。转录因子MYB、AP2/ERFERF、NAC、GARP-G2-like、HB-HD-ZIP和WRKY与单萜合成基因相关性较强，且6类转录因子之间存在较强相关性，其可能通过互作调控果实单萜合成或其他成熟衰老过程。     

1-MCP对‘绿宝石’梨采后成熟衰老期间H2O2含量及其相关酶活性的影响

为探讨1-甲基环丙烯(1-MCP)对梨果实采后成熟衰老过程中过氧化氢(H2O2)代谢的作用,试验以‘绿宝石’梨为材料,用1.0 μL/L 1-MCP密封处理24 h,空气密封为对照,研究1-MCP对‘绿宝石’梨室温(25±2)℃贮藏过程中果实硬度、可溶性同形物含量(SSC)、呼吸速率、H2O2含量、抗氧化酶活性[超氧化...     

Effect of salicylic acid (SA) on delaying fruit senescence of Huang Kum pear

This experiment was undertaken to explore the effect of salicylic acid (SA) at different concentrations on regulating fruit senescence of Huang Kum pear. Through dipping fruits and fruit discs for a series of hours in SA solution, enzyme activities and physiological characteristics of Huang Kum pear were determined. The results revealed that SA enhanced the activity of superoxide dismutase (SOD) and peroxidase (POD) enzymes at 0.02 mmol/L and at 0.002 mmol/L with the treatment of dipping fruit discs for 4 h and 12 h, respectively. The malondialdehyde (MDA) contents were reduced at 0.002 mmol/L for 12 h, and water loss ratio was decreased at 0.5 mmol/L after 48 h of treatment. It was concluded that SA at lower concentrations could delay the senescence of Huang Kum pear fruit.     

金秋梨幼果期钙处理对果实衰老相关生理的影响

以金秋梨为试材,采用对比试验,研究了幼果喷钙对金秋梨果实冷藏期间生理特性的影响,分析了钙对金秋梨采后生理变化的调节机制。结果表明,钙处理对提高果实钙含量和硬度有明显效应,降低了贮藏期间发病率,显著抑制超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性的降低,显著降低果实膜透性和减少丙二醛（MDA）含量,从而降低果实贮藏期间膜脂过氧化作用,延缓果实后熟衰老。配施萘乙酸（NAA）可增强喷钙效果。     

Studies on Changes of Phenolics in the Apple Fruit Peel in Response to Light Intensity

Fruits from 9-year-old apple trees (Malus domestic Borkh. cv. Fuji) were wrappedby two-layer bag in middle of June, and bags were removed in middle of September. The bag-removed treatment was performed in following three ways: once-removing all inner and outer bags; twice-removing bag (removing inner bag 3 d after removing outer bag); dipping 5 s in 1 mM salicylic acid (SA)after once-removing all bags. Changes of phenolic compounds in fruit peel in response to light environment were studied before or after removing bag. The results showed that the bagged treatment could significantly decrease the contents of UV (ultraviolet) absorbing compounds,rutin and anthocyanins of the fruit peel, but increased chlorogenic acid level. After removal of bag, the level of SA and quercetin in the fruit peel appeared peak 1 d after bag-removed,except that quercetin was decreased in SA-treated fruits. UVabsorbing compounds, anthocyanins and rutin were kept increasing all along after the removal of bag, except that chlorogenic acid was decreased during being retained inner bag in the treatment of twice removing bag. The positive correlations existed between changes of UV absorbing compounds and changes of anthocyanins or rutin. It was suggested that light played an important role in phenolics metabolism.     

褪黑素处理对梨果实采后黑斑病及贮藏品质的影响

[目的]探究褪黑素(Melatonine,MT)处理对梨果实采后黑斑病及贮藏品质的影响,为外源物质调控果实抗采后病害及贮藏品质提供理论依据和参考.[方法]以'翠冠'梨果实为试验材料,喷施0.1 mmol·L-1 MT溶液置室温,48 h后沿梨果实赤道两侧刺直径1 mm、深度3 mm大小两个小孔,待伤口晾干后注入20μL...     

枣果实不同发育阶段蛋白质组动态研究

枣果实在发育和成熟过程中涉及多种复杂的生理生化变化。为了解枣果实在发育过程中蛋白质组的变化情况,利用液质联用(LC-MS)技术对主栽制干枣品种——骏枣幼果期、膨大期、白熟期、初红期和全红期5个阶段的果实进行了非标记定量蛋白质组动态变化研究。结果表明,各时期果实与其前一阶段相比分别有差异表达蛋白189、356、82、69个。基因本体(GO)注释结果显示:幼果与膨大果的差异蛋白主要富集在核酸代谢过程,膨大期果实与白熟果的差异蛋白主要富集在碳水化合物代谢过程。从5个阶段的果实中分别鉴定到604、186、91、83、189个特异表达蛋白。Mercator注释和KEGG通路注释结果显示,幼果期特异表达蛋白主要参与了RNA转运通路。加权基因共表达网络(WGCNA)分析显示,幼果期高表达蛋白主要与核糖体合成相关。总之,枣果实在白熟期以前与细胞分裂和膨大相关的蛋白表达量和富集程度高,从白熟期开始与碳水化合物积累相关的蛋白质表达上调。本研究从蛋白质组学水平阐明了枣果实发育成熟过程中蛋白质组分的动态变化,为提高枣果实品质奠定了理论基础。     

苹果果皮酚类物质对光强变化的适应性研究

 以套袋3个月的富士苹果为试材,研究光强变化对果皮酚类物质的影响。结果表明,未套袋果实果皮的UV吸收物、芦丁、花青苷含量明显高于套袋果实的,而绿原酸含量降低。套袋果实除袋后,果皮绿原酸、UV吸收物、花青苷、芦丁呈现上升趋势;摘袋后用水杨酸(SA)浸蘸处理提高了果皮SA、槲皮素含量。     

海马齿根系响应盐胁迫的转录组分析

【目的】对盐胁迫下海马齿根系进行转录组测序分析,挖掘海马齿根系耐盐相关基因,为揭示海马齿耐盐的分子机制提供参考。【方法】利用Illumina测序技术对0 mmol/L NaCl （对照组）和400 mmol/L NaCl胁迫处理（盐胁迫处理组）下的海马齿根系进行转录组测序分析,从中筛选出差异表达基因,选取13个基因进行实时荧光定量PCR （qRTPCR）检测,以验证转录组数据的可靠性。【结果】在海马齿根系转录组中共鉴定出305145个转录本,平均长度为622 bp,其中,对照组有146177个长度>300 bp的转录本,盐胁迫处理组有72173个长度>300 bp的转录本;共有65535条Unigenes在Nr、GO、Swiss-Prot、COG和KEGG五大数据库注释成功,占Unigenes总数的52.36%。对照组和盐胁迫处理组共有65535个差异Unigenes,其中,有182个热休克蛋白基因。对照组和盐胁迫处理组间共有24042个差异表达基因,从中选取13个基因进行qRT-PCR检测,结果显示,9个基因表达上调,其余4个基因表达下调,与转录组测序结果一致。24042个差异表达基因中,共有10106个显著差异基因富集到129条代谢通路,其中富集程度排名前10的代谢途径为核糖体、次级代谢生物合成、RNA转运、内吞作用、剪接体、甘油磷脂代谢、内质网加工、吞噬、醚脂类代谢和植物-病原体相互作用,参与盐胁迫相关的硫代谢、脯氨酸积累、活性氧（ROS）代谢、与盐胁迫相关的钙信号通路和过氧化氢代谢等途径的差异基因上调。【结论】在盐胁迫下海马齿差异表达基因如小分子量热激蛋白基因、抗氧化酶相关基因及与离子交换相关基因发挥了重要调控作用。     

CaCl_2处理对采后柿果实生理变化的影响

以延津牛心柿为试材,对采收后的果实采用不同浓度的CaCl2处理,然后测定其软化衰老过程中果实硬度、可溶性固形物含量以及单宁含量的动态变化情况,结果表明:采后的柿果实用6%CaCl2处理能有效抑制其软化进程和单宁含量的降低,有效地延缓果实后熟和衰老进程。     

钙对苹果果实质膜脂肪酸组成及过氧化作用的影响

采用对比试验,研究了幼果喷钙对苹果果实质膜脂肪酸组成及过氧化作用的影响.结果表明:喷钙能显著增加棕榈酸百分比和降低亚油酸百分比,显著降低果实质膜透性、增加SOD活性和减少MDA含量,从而降低贮藏期间果实脂肪酸不饱和度和质膜透性及膜脂过氧化作用,延缓果实后熟衰老.配施NAA可增强喷钙效果.     

利用RNA-Seq鉴定甘蓝型油菜叶片干旱胁迫应答基因

【目的】利用RNA Sequencing（RNA-Seq）技术比较2种不同生长条件下甘蓝型油菜苗期叶片转录组,鉴定油菜叶片干旱胁迫应答相关基因,从转录组水平揭示油菜适应干旱胁迫环境的分子机制。【方法】提取正常生长（ZY）和自然失水处理（ZY8D）的六叶期甘蓝型油菜中油821的叶片总RNA,以Illumina Hiseq 2000平台进行RNA-Seq分析。利用NGSQCTookit v2.3.3去除低质量和包含模糊碱基的reads。以甘蓝型油菜亲本物种白菜染色体v1.5和甘蓝Scaffold v1.0为参考序列,采用TopHat2-Cufflinks-Cuffmerge-Cuffdiff标准流程进行差异表达基因（differential expressed genes,DEGs）筛选。对上调和下调DEGs分别采用Cytoscape v3.1.0中的BiNGO和KOBAS2.0进行基因本体（gene ontology,GO）和京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes,KEGG）代谢途径富集分析。选择上调和下调DEGs各3个,以实时荧光定量PCR（quantitative real-time PCR,qRT-PCR）验证RNA-Seq结果的可靠性。【结果】过滤低质量reads后,ZY和ZY8D分别保留了26 192 312和28 378 899对高质量reads用于DEGs筛选,其中86.6%和85.8%的reads能准确比对到参考序列上,说明RNA-Seq结果和参考序列可靠。DEGs鉴定结果表明3 657个基因受干旱胁迫诱导差异表达,其中上调表达基因1 431个,下调表达基因2 226个。GO富集分析发现上调表达基因主要与非生物胁迫响应和化学刺激响应相关,其中,参与水分胁迫响应和脱落酸（abscisic acid,ABA）刺激响应的基因分别有127和141个,而下调表达基因与植物病原菌防御、蛋白激酶活性和水杨酸（salicylic acid,SA）刺激相关。KEGG富集分析表明上调表达基因主要富集于苯丙烷和类胡萝卜素的生物合成及淀粉与蔗糖代谢途径,而下调表达基因主要富集于植物-病原菌互作和植物激素ABA、SA和茉莉酸（jasmonic acid,JA）信号转导途径。qRT-PCR检测6个DEGs的表达模式与RNA-Seq分析结果一致,证实了RNA-Seq结果的可靠性。【结论】RNA-Seq分析鉴定出3 657个甘蓝型油菜叶片干旱胁迫应答基因。GO和KEGG代谢途径分析明确了差异表达基因富集的分子功能与代谢途径。     

Changes of Activities in NAD Kinase and NADP Phosphatase During Ripening and Senescence of Tomato and 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 frnits 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.     

两种1-MCP制剂对丰水梨的贮藏效果

对自然室温条件下两种1—MCP制剂（EthylBloc和Ancip）对丰水梨果实贮藏效果的影响进行了研究．结果表明，两种处理都可以延缓果实硬度的下降，维持果实贮藏期间较高的可溶性固形物（TSS）含量和好果率，降低果实质量损失，使果实的风味和品质优于同期对照果实．两种处理都可以通过降低果实丙二醛（MDA）含量来延缓其衰老．其中EthylBloc处理贮藏效果较好，其果实货架期可比对照延长5d以上；Ancip处理果实的货架期比对照延长时间不超过5d．     

1-MCP对库尔勒香梨采后活性氧相关代谢的影响

[目的]研究1-甲基环丙烯(1-Methylcyclopropene,1-MCP)处理延缓库尔勒香梨采后衰老的机理,为香梨保鲜提供理论依据.[方法]以库尔勒香梨(Pyrus sinkiangensis Yu)为试材,研究1-甲基环丙烯处理对香梨果实贮藏期间活性氧代谢相关酶活性、超氧阴离子(O2·-)、丙二醛(MDA)、果皮色调角(h°)的影响.[结果]贮藏前期,1-MCP处理抑制了香梨果实超氧化物歧化酶(SOD)、过氧化物酶(POD)的活性,后期提高了POD的活性.整个贮藏期过氧化氢酶(CAT)活性低于对照;1-MCP处理果实丙二醛(MDA)含量低于对照果实;1-MCP处理在贮藏前期抑制超氧阴离子的产生,保持了较高的果实色调角.[结论]1-甲基环丙烯处理能够有效降低香梨活性氧产生速率,延缓果实衰老.     

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.     

不同成熟度杏果实采后抗坏血酸和超氧自由基代谢的变化

本文以两种成熟度的杏果实为材料,研究了杏果实采后膜脂过氧化与果实成熟衰老的关系。结果表明:两种成熟度杏果的膜脂过氧化特征不同,且采后均具有叶绿素和抗坏血酸降解的特征,绿熟果实中叶绿素和抗坏血酸降解与(?)产生速率升高呈显著的正相关。绿熟杏果实采后(?)产生速率持续升高,SOD活性和丙二醛含量先是快速增加而后下降;黄熟杏果实采后(?)产生速率、SOD活性和丙二醛含量则呈下降趋势。说明超氧自由基代谢失衡,导致膜脂过氧化加剧是果实成熟哀老的原因之一。