ABA和乙烯互作调控葡萄VlMybA1和VlMybA2表达并促进果皮着色

以‘巨峰’葡萄(Vitis vinifera×V. labrusca‘Kyoho’)为试材，研究了不同发育阶段果实花青素和ABA积累和乙烯释放的变化模式，及其与其合成或调控关键基因表达量的变化。基因表达、启动子响应和相对丰度分析表明，外源ABA处理上调了VlMybA1和VlMybA2表达，其中VlMybA1显著表达，促进果皮着色；外源ACC处理显著上调了Vl Myb A2表达，促进果皮着色。ABA和乙烯存在互作，100μmol·L^-1的外源ABA处理能够大幅度诱导内源乙烯的释放，500μmol·L^-1的外源ACC处理能够显著提高内源A BA含量；外源ABA处理能通过提高果皮内源乙烯释放水平增加VlMybA2的相对丰度。总之，ABA能直接调控VlMybA1表达，通过乙烯间接调控VlMybA2表达，促进葡萄果皮着色。     

脱落酸对桃果实成熟软化和乙烯生物合成的影响

以"白丽"桃为试材,分析了果实室温条件下贮藏过程中外源ABA(脱落酸)处理和NDGA(去甲二氢化愈创木酸,ABA生物合成抑制剂)处理后多聚半乳糖醛酸酶、果胶甲酯酶、ACC合成酶和ACC氧化酶含量的变化,探讨了在室温贮藏条件下ABA对果实成熟软化和乙烯生物合成的影响。结果表明:外源ABA处理可以提高多聚半乳糖醛酸酶和果胶甲酯酶的活性,加速果实硬度软化进程。外源ABA处理可以提高ACC合成酶和ACC氧化酶的活性,使果实乙烯释放量增加并且乙烯释放高峰提前出现。而NDGA处理则可以抑制多聚半乳糖醛酸酶的活性,果胶甲酯酶活性与清水相比虽然有降低,但抑制效果不显著。NDGA对果实硬度下降也有延缓作用。NDGA可以抑制ACC合成酶和ACC氧化酶的活性,从而降低乙烯释放量并且使乙烯释放高峰延时出现。     

安哥诺李果皮花青苷与内源激素、酶活性变化规律及其相关性

为了明确李果实花青苷合成的调控因子,以安哥诺李为试材,对果皮花青苷、内源激素和PAL、PPO活性的变化以及内源激素、酶活性与花青苷的相关性进行研究。结果表明,果皮花青苷含量在发育前期含量很低,发育后期含量迅速升高,呈上升趋势;果皮中ABA、乙烯、ZT含量和PPO活性与花青苷的变化趋势相同,乙烯释放量与花青苷呈极显著正相关关系r=0．8885**,ABA和ZT含量与花青苷呈显著正相关关系r=0．7159*和r=0．7476*,PPO活性未达显著水平;果皮中IAA、GA3含量和PAL活性变化在发育前期含量较高,在发育后期含量较低,果实发育过程中IAA、GA3含量呈逐渐下降趋势,与花青苷呈显著负相关关系r=-0．7039*和r=-0．8238*,PAL活性未达显著水平。     

ABA对‘巨峰’葡萄采后成熟关键基因表达的影响

利用ABA、NDGA(Nordihydroguaiaretictic acid,NCED酶抑制剂)和氟啶酮(Fluridone)、乙烯利和AS6(3’-hexylsulfanyl-ABA,PYL-PP2C受体拮抗剂)处理‘巨峰’葡萄采后果实,分析果实硬度、花色苷、芳香挥发物及相关基因表达量等的变化。结果表明,ABA可促进花色苷、可溶性固形物和香气挥发物形成,降低可滴定酸含量与果实硬度。NDGA、氟啶酮、AS6的效果与ABA相反。VvMYBA2、VvUFGT和VvEGS1的表达可促进花色苷的积累和芳香挥发物的形成,而Vv LAR1则减少花色苷积累。施加外源ABA,可促进果实内源ABA含量增加和VvOPR3、VvACO1和VvSUT11的表达;AS6处理也可增加内源性ABA含量,而NDGA、氟啶酮则相反。VvNCED1、VvCYP707A1、VvPYL8的表达与内源ABA含量基本一致,相比之下,AS6可下调V PYL8表达量。VvPP2C49的表达变化与内源ABA相反,AS6促进VvPP2C49表达。这些结果表明果实内源ABA含量影响花色苷、香气挥发产物的形成,促进果实软化和糖代谢。     

甜樱桃果实NCED基因的克隆及其表达

为了进一步研究ABA在甜樱桃果实成熟过程中的作用,通过RT-PCR以及RACE-PCR方法从甜樱桃果实克隆得到了ABA合成关键酶NCED基因片段PacNCED1及其3′ 末端序列,从乙烯利处理果实中克隆得到了乙烯合成关键酶ACO基因片段PacACO1。推测的PacNCED1和PacACO1氨基酸序列与其它物种的NCEDs和ACOs氨基酸序列具有很高的同源性。通过半定量RT-PCR及定量RT-PCR方法分析了PacNCED1和PacACO1的表达模式,发现PacNCED1在甜樱桃果实发育的整个过程以及不同部位都有表达,在果肉和种子中的表达不断增加,在果实成熟之前达到最高峰,而在果柄中的表达则在果实完熟时达到最大。PacNCED1在未失水的叶片和根中有微弱表达,但在失水叶片中的表达急剧上升,表现出与失水的相关性。外源ABA和乙烯利能促进PacNCED1在果实中的表达,而NDGA和IAA对PacNCED1在果实中的表达没有显著影响。在甜樱桃果实发育过程中并没有检测到PacACO1的表达信号,但是外源乙烯利和ABA处理能诱导其在果实中的表达。     

脱落酸和2,4-表油菜素内酯对葡萄成熟过程中果实内源激素含量的影响

以酿酒葡萄"赤霞珠"和"烟73"为试材,研究了脱落酸(ABA)、2,4-表油菜素内酯(EBR)和BR生物合成抑制剂油菜素唑(Brz)处理对葡萄果实乙烯生成量、内源生长素(IAA)和赤酶素(GA3)含量的影响。结果表明:外源ABA和EBR处理促进了果实乙烯的释放,同时促进了果实着色,ABA和EBR处理后果实内源IAA和GA3含量略低于对照,但总体无显著差异。在不同浓度EBR处理中,以0.4mg/LEBR处理效果最显著。Brz处理抑制了乙烯的释放,推迟了果实着色,但对果实内源IAA和GA3含量没有显著影响。     

甜樱桃红灯果实发育过程中果肉及种子内源激素含量变化动态

以9a生红灯甜樱桃(Prunus avium L.)为试材,对其果实不同发育时期果肉及种子中内源激素含量的变化动态进行了测定分析,结果表明,甜樱桃果肉及种子中内源激素含量与其果实的生长发育有密切关系。果实发育第Ⅰ速长期(盛花后5～15d),果肉中生长促进型激素(ZRS、IAA和GAS)含量均较高,进入硬核期(盛花后15～25d)均呈下降趋势,其中IAA和GAS均降到最低值,随果实的第Ⅱ次速长果肉中ZRS、IAA和GAS含量均迅速增加且分别出现最大值;与之相反,种子中ZRS、IAA和GAS含量在第Ⅰ、Ⅱ速长期较低,在硬核期含量最高,这与甜樱桃果实及种子的生长规律相吻合。果肉中ABA含量分别在盛花后5、15和35d出现高峰,与甜樱桃的落果(花)时期基本一致。     

早蜜梨果实生长发育期间内源激素含量变化的研究

采用高效液相色谱(HPLC)技术,测定了早蜜梨果实生长发育期间内源生长素(IAA)、赤霉素(GA3)、玉米素(ZT)和脱落酸(ABA)含量及其比值的变化,并分析了各激素与果实发育的关系.结果表明:果肉和种子中IAA含量分别在花后50 d和20 d出现高峰;GA3含量分别在花后20 d和60 d达到峰值;ZT含量的高峰则在花后40 d,而ABA含量分别在花后80 d和90 d最高,随后均呈逐渐下降.IAA/ABA、GA3/ABA和ZT/ABA比值对果实的细胞分裂与膨大起着重要调节作用.     

外源ABA和EBR处理对酿酒葡萄“赤霞珠”生长成熟的调控及内源激素的影响

以酿酒葡萄品种"赤霞珠"为试材,研究了外源脱落酸(Abscisic acid,ABA)和表油菜素内酯(Epibrassinolide,EBR)处理对其生长过程中不同组织(果肉、果皮、种子)内源激素的影响及其对葡萄生长成熟的调节作用。在转色前2周(花后52d)对葡萄进行外源ABA和EBR处理,采用高效液相色谱与质谱联用(HPLC-MS)技术测定ABA和EBR处理后不同时期葡萄果肉、果皮、种子中ABA、生长素(Auxin,IAA)、赤霉素(Gibberellin,GA_3)和水杨酸(Salicylic acid,SA)的含量。结果表明:在葡萄果实生长发育过程中,外源添加ABA和EBR均能提高葡萄果实中还原性糖的含量,并可显著降低可滴定酸的含量,但不影响成熟时期还原糖的最终含量;在处理早期,ABA处理和EBR处理均能显著增加果肉中IAA、GA_3和SA的含量,说明2种处理能加速葡萄的发育,促进果实的膨大;ABA处理和EBR处理均能显著提高转色期葡萄果实ABA的含量,提前抑制种子中生长素运输到果实其它部位的通路,说明外源添加ABA和EBR均能促进葡萄成熟。试验表明,外源ABA和EBR处理均能加速葡萄果实的发育,促进果实的成熟。     

喷施GA3和乙烯利对柿果实成熟及内源GAS活性,ABA含量的影响

选用柿极晚熟品种宫崎无核,于果实发育第二期末(10月19日),叶面喷施100ppm的GA_3明显地抑制了果实的肥大生长、着色、糖的积累及果肉软化。对果实内源GAs活性和ABA含量进行分析的结果表明,叶面喷施GA_3使内源GAs活性大大提高,同时使ABA的增加受到抑制。与GA_3处理相反,果实发育第三期(10月1日),对中熟品种富有进行25ppm乙烯利的叶面喷施处理,明显地促进了果实的成熟,其作用尤以促进着色、加速果实软化最为明显。对内源GAs活性及ABA含量亦有明显影响,前者因乙烯利处理而降低,后者则因乙烯利处理而增大。     

ABA 和乙烯对荔枝果实成熟和着色的调控

 荔枝果实幼果期ABA 含量很高, 之后下降, 在转红前1 周急剧上升, 形成一个峰值。果皮、果肉乙烯释放量的变化与ABA 相似, 但其跃变发生在ABA 之后, 且跃变较小。伴随着乙烯的跃变, 有一个糖代谢和转化酶活性的跃变。于转熟前用STS 处理荔枝果实, 导致果实转色和成熟延迟。果实发育后期, ABA 急剧增加可能诱导了乙烯的合成, 乙烯启动果实成熟, 果皮转红着色。     

Effects of 1-aminocyclopropane carboxylic acid on the rate of ethylene release from detached fruiting spurs and on fruit abscission in apple

Summary

1-Aminocyclopropane carboxylic acid (ACC) was applied to apple (Malus × domestica Borkh.) trees or to individual fruiting spurs between full bloom (FB) and 31 d after full bloom (DAFB; mean fruit diameter of 20 mm) to determine the effects of ACC on the ethylene released from detached spurs and on fruit abscission. The rate of ethylene release from detached fruiting spurs increased 1 d after applying 50 mg l^–1 ACC at full bloom or 16 DAFB (mean fruit diameter of 10 mm), but ACC had a reduced effect on the rate of ethylene release when it was applied 31 DAFB (mean fruit diameter of 20 mm). ACC reduced fruit set in ‘GoldRush’ apple in a concentration-dependent manner in 2009 and in 2010, but had no thinning activity on ‘Pink Lady’ in 2010. ACC at 200 mg l^–1 resulted in the yellowing and abscission of ‘GoldRush’ spur leaves in 2009, but not in 2010. The effects of combining ACC and 5 mg l^–1 naphthaleneacetic acid on fruit set in ‘GoldRush’ apple were additive. The application of 200 mg l^–1 ACC when fruit are ca. 10 mm in diameter may therefore be a useful chemical thinning treatment for apple fruit.     

Oxylipin affects ethylene metabolism and ethylene receptor gene expression levels in peach fruit (Prunus persica L. Batsch)

The effects of two oxylipins – jasmonic acid (JA) and 9, 10-ketol-octadecadienoic acid (KODA) – on ethylene metabolism and ethylene receptor genes were examined in peach fruit (Prunus persica L.) harvested at 88 days after full bloom (DAFB), in the pre-climacteric stage, and at 102 DAFB, at the beginning of the climacteric stage. Immediate post-harvest applications of either n-propyl dihydrojasmonate (PDJ) (a JA analog) or KODA stimulated ethylene production and 1-aminocyclopropane-1-carboxylate (ACC) concentrations in fruit that were harvested at 88 and 102 DAFB. KODA application significantly increased levels of ACC oxidase (PpACO1) expression at 5 and 8 days after treatment (DAT) in 88-DAFB-harvested fruit, and at 1 DAT in 102-DAFB-harvested fruit. Expression levels of ethylene response factors (PpERS1) were not influenced by either PDJ or KODA applications at 88 or 102 DAFB. In contrast, levels of ethylene-receptor-coding genes (PpETR1) expression in 88-DAFB KODA-treated fruit increased at 8 and 14 DAT. Expression levels of constitutive triple response (PpCTR1) in the untreated control fruit decreased with DAT in both 88- and 102-DAFB-harvested fruit.

These results suggest that certain oxylipins may promote ethylene production in peach fruit, and that PpETR1 and PpCTR1 may be associated with the autocatalytic system of ethylene production.     

鸭梨,茌梨果实采前乙烯生物合成特性研究

鸭梨、茌梨果实均从盛花后１４５天进入呼吸跃变，同时伴随着乙烯释放高峰的出现。鸭梨的乙烯合成酶（ＥＦＥ）活性和氨基环丙烷羧酸（ＡＣＣ）含量高于茌梨，乙烯释放量的峰值是茌梨的４倍，两品种果实的ＥＦＥ活性变化与果实的呼吸跃变和乙烯释放量的变化同步，ＡＣＣ含量自盛花后１３５天，一直呈止升趋势。两品种果实内含有亚精胺、腐胺和精胺三种内源多胺，亚精胺与梨果实乙烯生物合成有关。     

Effects of ethylene,abscisic acid and auxin on fruit abscission in water dropwort (Oenanthe stolonifera DC.)

Fruit abscission in water dropwort (Oenanthe stolonifera DC.) was investigated by measuring fruit removal force (FRF) affected by fruit ages and plant hormones. FRF decreased rapidly 30 days after anthesis (DAA). Ethylene evolution increased rapidly at 35 DAA and ABA concentration increased gradually after 20 DAA. ACC and ethephon decreased the FRF of fruit explants and the promoting effect of ACC was delayed by silver thiosulphate and cycloheximide, but not by norbornadiene and actinomycin D. ABA enhanced fruit abscission without increasing ethylene evolution. However, the observation that CoCl₂ delayed abscission induced by ABA indicates possible involvement of ethylene in the ABA effect. Treatment with NAA, fenoprop and IAA had no effect on 30 DAA fruit explants, but IAA delayed abscission of 25 DAA explants. This suggests that the effect of auxins may differ with fruit age when used as control agents.     

油菜素内酯和脱落酸调控葡萄果实花色苷合成的研究

【目的】为研究2,4-表油菜素内酯(2,4-epibrassinolide,EBR)和脱落酸(abscisic acid,ABA)处理对葡萄花色苷合成与可溶性固形物含量、果皮苯丙氨酸解氨酶(PAL)和类黄酮糖基转移酶(UFGT)活性以及果实内源ABA含量的影响,探索油菜素内酯调控葡萄果实成熟及花色苷合成的机理。【方法】以酿酒葡萄‘赤霞珠’(Cabernet Sauvignon)和‘烟73’(Yan 73)为试材,在葡萄转色前分别用0.1、0.4、0.8 mg.L-1EBR,1mg.L-1Brz(brassinazole,BR生物合成抑制剂)和200 mg.L-1ABA,均匀喷施于葡萄果实,在葡萄成熟过程中测定葡萄果皮花色苷含量及PAL和UFGT酶活性,同时测定果实ABA和可溶性固形物含量。【结果】在果实着色初期,‘赤霞珠’和‘烟73’葡萄果皮PAL和UFGT活性及果实内源ABA含量均逐渐升高,当果实接近成熟花色苷含量基本稳定时ABA含量开始降低。与对照相比,0.4mg.L-1EBR和200 mg.L-1ABA处理显著增加了果实内源ABA含量,提高了果皮PAL和UFGT活性,促进了果皮花色苷的合成和果实可溶性固形物的积累。0.1 mg.L-1和0.8 mg.L-1EBR处理总体增加了果实内源ABA含量,促进了花色苷的合成和可溶性固形物的积累,并提高了UFGT和PAL酶活性,但比0.4 mg.L-1EBR处理提高的幅度小,且差异显著。1 mg.L-1Brz处理使果实ABA的合成推迟,果实可溶性固形物含量、果皮PAL和UFGT活性以及花色苷含量均低于对照,但差异不显著。【结论】外源EBR和ABA处理促进了葡萄成熟和花色苷合成,在不同浓度EBR处理中,以0.4 mg.L-1处理效果较好;内源ABA可能参与了EBR对葡萄成熟和花色苷合成的调控。     

苹果采前落果与内源激素的关系

为了探讨苹果采前落果与内源激素之间的关系,在采前8周间定期测定了不同苹果品种的果柄、果台和离层形成部位组织中IAA、ABA含量;在采收前20d内定期测定了离层部位组织中细胞壁分解酶(Cellulase)的活性;在收获期测定了果实乙烯的发生量。结果表明:不同品种果柄、果台和离层部位组织中IAA和ABA含量变化有差异,但它们变化的总趋势相似,都是随着果实成熟IAA含量下降,而ABA含量上升;不同品种的成熟果实中乙烯发生量有很大差异,以落果多的品种显著大于落果少的品种;采前落果重的品种离层部位组织中细胞壁分解酶的活性在果实成熟期急剧增加。由于果实进入成熟阶段后,IAA含量下降,ABA含量升高,ABA/IAA之间的相对平衡被打破,高ABA/IAA以及高乙烯会刺激离层组织中细胞壁分解酶的活性增高,进而促进离层形成,这可能是导致落果发生的原因。     

Effects of abscisic acid agonist or antagonist applications on aroma volatiles and anthocyanin biosynthesis in grape berries

The effects of applying abscisic acid (ABA), Abz (abscinazole, an inhibitor of CYP707A), and nordihydroguaiaretic acid ((NDGA), an inhibitor of 9-cis-epoxycarotenoid dioxygenase (NCED)) on fruit firmness, anthocyanin biosynthesis, aroma volatiles, and their related genes were investigated in grape berries (Vitis vinifera L. × Vitis labrusca L. cv. Kyoho). ABA and Abz application promoted the accumulation of anthocyanin and aroma volatiles, the activity of polygalacturonase (PG), pectin methylesterase, and cellulase, but reduced fruit firmness. On the other hand, NDGA showed the opposite tendencies. Abz decreased VvCYP707A1 expression but increased VvPYL8 expression. VvPP2C49 expression was negatively correlated with ABA concentration. The expression levels of VvMYBA2 and VvUFGT correlated with anthocyanin concentration, unlike those of VvLAR1. ABA and Abz application increased endogenous ABA concentrations and the expression levels of VvOPR3, VvACO1, and VvPG, whereas NDGA application decreased them. These results suggest that ABA concentrations influence anthocyanin formation and fruit softening through the expression of VvMYBA2, VvUFGT, and VvPG.