香蕉一个Ⅲ类酸性几丁质酶基因与果实成熟关系的研究

为了解Ⅲ类酸性几丁质酶基因(MaCHⅢ)与香蕉果实采后成熟过程的相互关系,对经乙烯和1-甲基环丙烯(1-MCP)处理的巴西香蕉果实采后乙烯释放量、Ⅲ类酸性几丁质酶基因(MaCHⅢ)表达以及几丁质酶活性进行了测定.结果显示:(1)乙烯催熟处理的香蕉果实,乙烯释放量比对照处理的果实提前15 d达到高峰;1-MCP处理的香蕉果实,乙烯生物合成和果实成熟明显受到了抑制.(2)外源乙烯加速了MaCHⅢ基因的下调表达和Ⅲ类酸性几丁质酶活性的下降,MaCHⅢ表达量和Ⅲ类酸性几丁质酶活性分别在采后第3天和第4天下降到最小值.(3)1-MCP处理使MaCHⅢ基因呈现上调表达,Ⅲ类酸性几丁质酶活性上升,MaCHⅢ基因表达量和Ⅲ类酸性几丁质酶活性分别在采后18 d和25 d达到高峰.研究表明,MaCHⅢ基因可能与香蕉果实采后成熟呈负相关.     

1-MCP对香蕉果实货架期的影响

香蕉果实经乙烯利处理后当天和1d分别用1-甲基环丙烯(1-MCP)处理,果实色泽转变、软化及淀粉降解均受到明显抑制,货架期延长5d以上;而乙烯利处理后贮放2d或3d再用1-MCP处理已失去对果实后熟的抑制作用。香蕉果实经1-MCP处理后在常温下贮藏11d完全恢复对乙烯敏感。     

香蕉MaBTB基因的表达分析及亚细胞定位

利用荧光定量PCR分析在不同处理下的香蕉采后果实中MaBTB基因的表达,在正常成熟的果实中,MaBTB基因的表达与乙烯的释放量呈正相关;相反,在1-MCP处理的香蕉果实中,该基因的表达相对变化不明显;用乙烯处理的香蕉果实,其表达量在第3天达到高峰,比正常成熟的早11 d。这些结果表明MaBTB基因在香蕉采后果实中是受乙烯诱导表达的。最后,亚细胞定位分析表明MaBTB基因定位在细胞核上。     

1-甲基环丙烯对低温贮藏的香蕉果实后熟的影响

香蕉果实先在6℃下贮藏不同时间后转入常温用1-甲基环丙烯(1-MCP)处理或先以1-MCP处理后转入6℃下贮藏不同时间,然后在常温下用乙烯利处理,结果表明, 经1-MCP处理过的果实对乙烯不敏感,后熟过程受抑;而未经1-MCP处理的果实能够后熟,并有明显的呼吸及乙烯峰出现.     

1-甲基环丙烯处理对采后杨桃果实软化和细胞壁代谢的影响

为探讨1-甲基环丙烯(1-methylcyclopropene, 1-MCP)延缓采后杨桃果实软化的作用机理,本文研究了0.6 μL/L 1-MCP处理对在(15±1)℃、相对湿度90%下贮藏的‘香蜜’甜杨桃(Averrhoa carambola Linn. cv. Xiangmi)果实软化和细胞壁代谢的影响。结果表明：与对照果实相比,1-MCP处理可保持较高的杨桃果实硬度,有效降低果胶酯酶(pectinesterase, PE)、多聚半乳糖醛酸酶(polygalacturonase, PG)、纤维素酶等细胞壁降解酶活性,延缓原果胶、纤维素、半纤维素含量的下降和水溶性果胶含量的增加。因此认为,0.6 μL/L 1-MCP处理能有效控制采后‘香蜜’甜杨桃果实的软化进程,延长果实保鲜期。     

1-MCP对梨采后某些生理生化指标的影响

1.0μL·L-1 1-MCP抑制梨品种"京白"、"五九香"、"锦香"果实采后呼吸速率.随着冷藏期的延长,货架期间经1-MCP处理的果实呼吸速率逐渐上升;冷藏后货架期间1-MCP延缓果实硬度下降,梨果实的酸度、淀粉、果皮叶绿素含量得以保持,但作用大小因品种而异;1-MCP还可以抑制"锦香"梨冷藏和货架期间黑皮病的发生,防止果实腐烂.     

甜柿采后生理特性及对1-MCP处理的反应

以甜柿品种‘阳丰’为材料,在20℃和0℃贮藏条件下研究了1-MCP(1-甲基环丙烯)处理对不同成熟度甜柿果实采后乙烯释放速率、呼吸速率和品质特性的影响。结果表明:1-MCP处理可延缓贮藏和货架期间甜柿果实的软化、抑制呼吸速率和可溶性固形物含量(SSC)的变化,但对乙烯释放速率的作用不一致。1-MCP处理对成熟度I果实的效果优于成熟度Ⅱ。低温贮藏虽然能显著延缓果实硬度的下降,但在0℃贮藏30、60和90 d后7 d货架期结束时,对照果完全软化,而经1-MCP处理后果实果肉硬度仍保持“脆”性。因此,贮前0.50μL.L-11-MCP处理结合低温贮藏是延长‘阳丰’甜柿贮藏期的有效途径,具有广泛的应用前景。     

钙对香蕉采后乙烯释放的抑制

本实验用CaCl_2溶液对香蕉(Musa acuminata cf. 'Dwarf Davendish')组织进行真空浸透处理,研究Ca~(2 )对香蕉采后乙烯释放、EFE活性、ACC水平以及ACC/MACC比值的影响。结果表明,Ca~(2 )处理可抑制香蕉果皮和果肉组织乙烯生成,对抑制果皮的乙烯生成尤为明显。Ca~(2 )处理还可降低内源ACC水平,抑制EFE活性。结果还显示,Ca~(2 )处理对组织中ACC/MACC比值有一定影响。     

不同后熟期‘菊水’梨果实对外源乙烯和1-MCP处理的生理响应

用80uL·L-1外源乙烯和1.0 uL·L-11-甲基环丙烯(1-MCP)处理不同后熟期'菊水'梨果实,分析处理后果实品质和生理指标在(25±1)℃贮藏温度下的变化特征.结果显示:在采收当天(采后0 d)和呼吸跃变初期(采后4 d),外源乙烯处理能明显促进果实硬度和可溶性同形物含量(SSC)的下降,降低活性氧清除酶(SOD、CAT和APX)的活性,提高呼吸速率和乙烯释放速率,促进果实后熟,1-MCP处理却表现出与乙烯相反的效应,且采收当天比呼吸跃变初期的作用效果更明显;在呼吸跃变中期(采后12 d),外源乙烯和1-MCP处理效果均不明显.研究发现,外源乙烯能促进果实后熟而1-MCP却抑制果实后熟,其效果因处理果实后熟期的不同而存在显著差异,果实后熟程度越高,其处理的效果越不明显.     

CO2和1-MCP组合处理对磨盘柿贮藏效果的影响

以磨盘柿采后果实为材料,研究1-MCP的处理以及CO2脱涩与1-MCP组合处理对磨盘柿室温和0~1℃贮藏过程中果实硬度、可溶性单宁含量、总抗坏血酸(TAA)含量以及抗氧化活性的影响。结果显示:(1)贮藏过程中柿果实硬度随时间延长呈下降趋势,室温贮藏CO2处理5 d、CO2处理后进行1-MCP(CO2//1-MCP)处理10 d、1-MCP处理后进行CO2(1-MCP//CO2)处理15 d、CO2与1-MCP同时(CO2+1-MCP)处理30 d均完全软化,0~1℃贮藏75 d后硬度最高的是CO2+1-MCP处理(12.43 kg.cm-2),最低的是CO2//1-MCP处理(2.80 kg.cm-2),甚至低于CO2处理(5.71 kg.cm-2);(2)柿果实可溶性单宁和总抗坏血酸(TAA)含量与脱涩有关,CO2处理、CO2//1-MCP处理、1-MCP//CO2处理和CO2+1-MCP处理大幅低于CK和1-MCP处理,1-MCP处理抑制了可溶性单宁和TAA含量的下降;(3)室温贮藏中除CO2处理之外的处理柿果实总酚含量呈小幅上升趋势,0~1℃贮藏中CK、1-MCP处理、1-MCP//CO2处理和CO2+1-MCP处理的总酚含量稳定在9.91~12.38 mg.g-1FW之间,CO2处理和CO2//1-MCP处理于30 d之后迅速下降,至75 d时分别只有6.83和6.32 mg.g-1FW;(4)各组处理柿果实ABTS自由基清除能力和氧自由基清除能力值(ORAC)的变化趋势与总酚含量大致相当。研究发现,1-MCP能有效阻止磨盘柿果实贮藏期间的硬度、可溶性单宁含量、TAA含量以及抗氧化活性的下降,CO2脱涩与1-MCP处理的不同顺序对硬度和抗氧化活性影响巨大,但对可溶性单宁和TAA含量影响甚微;先CO2脱涩后1-MCP处理对磨盘柿贮藏效应影响不大,1-MCP处理和高浓度CO2脱涩同时进行是磨盘柿脱涩保鲜的最优方案。     

Ethylene rapidly up-regulates the activities of both monomeric GTP-binding proteins and protein kinase(s) in epicotyls of pea

It is demonstrated that, in etiolated pea (Pisum sativum) epicotyls, ethylene affects the activation of both monomeric GTP-binding proteins (monomeric G-proteins) and protein kinases. For monomeric G-proteins, the effect may be a rapid (2 min) and bimodal up-regulation, a transiently unimodal activation, or a transient down-regulation. Pretreatment with 1-methylcyclopropene abolishes the response to ethylene overall. Immunoprecipitation studies indicate that some of the monomeric G-proteins affected may be of the Rab class. Protein kinase activity is rapidly up-regulated by ethylene, the effect is inhibited by 1-methylcyclopropene, and the activation is bimodal. Immunoprecipitation indicates that the kinase(s) are of the MAP kinase ERK1 group. It is proposed that the data support the hypothesis that a transduction chain exists that is separate and antagonistic to that currently revealed by studies on Arabidopsis mutants.     

The effect of chemical structure on the antagonism by cyclopropenes of ethylene responses in banana

The compounds, cyclopropene, 1-methylcyclopropene, 3-methylcyclopropene, 1,3-dimethylcyclopropene, 3,3-dimethylcyclopropene, 1,3,3-trimethylcyclopropene, 3-methyl-3-vinylcyclopropene, and 3-methyl-3-ethynylcyclopropene, and 1,2-dimethylcyclopropene were tested as antagonists to the ethylene receptor in bananas. All of the compounds inactivated the receptor and the bananas did not respond to ethylene even at 1000 nL L^–1. Large differences were found in the concentration required (0.7–20,000 nL L^–1 for 24h) to inactivate the receptor and in the duration of inactivation (3–12 days at 24C depending on the compound). After this time the bananas responded to ethylene and appeared to ripen normally.     

1-甲基环丙烯对百合采后切花某些生理指标的影响

百合切花经1-甲基环丙烯(1-MCP)处理后瓶插寿命延长,花朵发育和衰老进程延缓,乙烯峰出现时间推迟.经1-MCP处理的亚洲百合的乙烯峰值和细胞膜透性降低,而麝香百合可溶性蛋白质含量则不受影响.     

1-甲基环丙烯(1-MCP)对油桃果实软化的影响

1-甲基环丙烯(1-MCP)可延缓油桃果实硬度的下降,阻止引起果实软化的细胞物质(淀粉、纤维素、果胶)的降解,抑制与果实软化相关的酶(淀粉酶、纤维素酶、多聚半乳糖醛酸酶)活性。     

Ligand-induced alterations in the phosphorylation state of ethylene receptors in tomato fruit

Perception of the plant hormone ethylene is essential to initiate and advance ripening of climacteric fruits. Since ethylene receptors negatively regulate signaling, the suppression is canceled upon ethylene binding, permitting responses including fruit ripening. Although receptors have autophosphorylation activity, the mechanism whereby signal transduction occurs has not been fully determined. Here we demonstrate that LeETR4, a critical receptor for tomato (Solanum lycopersicum) fruit ripening, is multiply phosphorylated in vivo and the phosphorylation level is dependent on ripening stage and ethylene action. Treatment of preclimacteric fruits with ethylene resulted in accumulation of LeETR4 with reduced phosphorylation whereas treatments of ripening fruits with ethylene antagonists, 1-methylcyclopropene and 2,5-norbornadiene, induced accumulation of the phosphorylated isotypes. A similar phosphorylation pattern was also observed for Never ripe, another ripening-related receptor. Alteration in the phosphorylation state of receptors is likely to be an initial response upon ethylene binding since treatments with ethylene and 1-methylcyclopropene rapidly influenced the LeETR4 phosphorylation state rather than protein abundance. The LeETR4 phosphorylation state closely paralleled ripening progress, suggesting that the phosphorylation state of receptors is implicated in ethylene signal output in tomato fruits. We provide insights into the nature of receptor on and off states.     

An early ethylene up-regulated gene encoding a calmodulin-binding protein involved in plant senescence and death

35S-Labeled calmodulin (CaM) was used to screen a tobacco anther cDNA library. A positive clone (NtER1) with high homology to an early ethylene-up-regulated gene (ER66) in tomato, and an Arabidopsis homolog was isolated and characterized. Based on the helical wheel projection, a 25-mer peptide corresponding to the predicted CaM-binding region of NtER1 (amino acids 796-820) was synthesized. The gel-mobility shift assay showed that the peptide formed a stable complex with CaM only in the presence of Ca(2+). CaM binds to NtER1 with high affinity (K(d) approximately 12 nm) in a calcium-dependent manner. Tobacco flowers at different stages of development were treated with ethylene or with 1-methylcyclopropene for 2 h before treating with ethylene. Northern analysis showed that the NtER1 was rapidly induced after 15 min of exposure to ethylene. However, the 2-h 1-methylcyclopropene treatment totally blocked NtER1 expression in flowers at all stages of development, suggesting that NtER1 is an early ethylene-up-regulated gene. The senescing leaves and petals had significantly increased NtER1 induction as compared with young leaves and petals, implying that NtER1 is developmentally regulated and acts as a trigger for senescence and death. This is the first documented evidence for the involvement of Ca(2+)/CaM-mediated signaling in ethylene action.