硝酸钙对嫁接网纹甜瓜果实糖含量及糖代谢相关酶活性的影响

为解决嫁接厚皮甜瓜含糖量下降的问题,以自根厚皮甜瓜和嫁接厚皮甜瓜为试材,探讨了不同浓度硝酸钙处理对嫁接厚皮甜瓜糖代谢的影响。结果显示：叶面喷施不同浓度硝酸钙对嫁接厚皮甜瓜果实糖含量的影响程度不同。与嫁接对照相比,0.5 %硝酸钙处理显著增加了嫁接厚皮甜瓜果实发育后期（处理后14～42 d）的果糖、葡萄糖、蔗糖含量,降低了棉籽糖和水苏糖含量,提高了肌醇半乳糖含量|0.2 %和0.8 %硝酸钙处理虽也在一定程度上影响着可溶性糖含量,但影响程度低于0.5 %硝酸钙处理|处理21 d后,3个浓度的硝酸钙处理均极显著增加了果实中总糖含量,且0.5 %硝酸钙处理还极显著增加了淀粉含量|但与自根对照相比,0.5 %硝酸钙处理的果糖、葡萄糖、蔗糖、淀粉和总糖含量仍未能增加至自根厚皮甜瓜水平。从果实的蔗糖代谢及半乳糖代谢相关酶来看,与嫁接对照相比,叶面喷施不同浓度的硝酸钙处理均显著或极显著的增加果实发育中期（处理后14～28 d）酸性转化酶、碱性α- 半乳糖苷酶和果实发育中后期（处理后28～42 d）蔗糖磷酸合成酶的活性,但对于中性转化酶和蔗糖合成酶活性影响不显著|与自根对照相比,0.5 %硝酸钙处理的酸性转化酶活性未达到自根对照水平,蔗糖磷酸合成酶活性大幅提升,与自根对照差异不显著。结果表明：与嫁接对照相比,叶面喷施不同浓度硝酸钙处理均可不同程度提高嫁接厚皮甜瓜果实淀粉、总糖、葡萄糖、果糖、蔗糖含量及其果实发育前中期的酸性转化酶、碱性α- 半乳糖苷酶和果实发育中后期的蔗糖磷酸合成酶等糖代谢相关酶活性,其中0.5 %硝酸钙处理效果最佳,增加了果实发育中后期蔗糖含量,具有改善嫁接厚皮甜瓜果实品质的作用。     

伴生小麦对黄瓜叶片衰老期间糖代谢相关指标的影响

采用田间试验研究伴生小麦栽培对黄瓜叶片衰老期间糖代谢相关指标的影响。结果表明,在黄瓜第4片叶叶龄为30、50 d时,伴生小麦处理的黄瓜叶片中可溶性糖、还原糖、葡萄糖和蔗糖含量均显著低于单作;在黄瓜第4片叶叶龄为30、40 d时,伴生小麦处理的黄瓜叶片中蔗糖合成酶与蔗糖磷酸合成酶活性均显著高于单作;伴生小麦处理的黄瓜叶片中性转化酶活性在第4片叶叶龄为20、60 d,酸性转化酶的活性在第4片叶叶龄为20、30、50 d时显著高于单作。综合分析糖代谢相关指标的变化,采用小麦伴生栽培模式,在一定程度上增强了黄瓜叶片糖代谢活动,有利于延缓叶片衰老。     

甜瓜肌醇半乳糖苷合成酶基因CmGAS1的表达与功能分析

甜瓜（Cucumis melo L.）是棉子糖系列寡糖（Raffinose Family Oligosaccharides,RFOs）转运型植物。肌醇半乳糖苷合成酶（GAS）是催化RFOs合成的关键酶。甜瓜CmGAS1基因（GenBank登录号为AY077642.1）的cDNA全长为1 903 bp,开放阅读框（ORF）为996 bp,编码331个氨基酸。CmGAS1蛋白分子量约为38 kD,理论等电点（pI）为4.81。氨基酸序列比对和系统进化树分析表明,CmGAS1氨基酸序列与黄瓜（AAO84915.1）亲缘关系最近,同源性为98.79%,与西瓜（Cla009222）同源性为97.58%。荧光定量结果表明,甜瓜叶片从幼叶（库）至成熟叶（源）CmGAS1的表达显著升高,第5节位叶片达到最大值。去掉50%叶片植株与对照相比,完全展开功能叶片的CmGAS1表达量明显升高。应用CmGAS1特异性启动子构建CmGAS1的过表达载体,并采用农杆菌介导进行甜瓜遗传转化,获得了PCR阳性转化植株,转化植株完全展开功能叶片的净光合速率以及叶片中的蔗糖、棉子糖、水苏糖含量与野生型相比均有所提高。推测CmGAS1在甜瓜叶片棉子糖系列寡糖合成过程中起重要作用。     

黄瓜叶片和韧皮部汁液中碳水化合物及相关酶的日变化

 以黄瓜叶片和韧皮部汁液为对象, 研究1个昼夜周期中各种碳水化合物含量及相关酶活性的变化。结果表明夜间至上午10: 00, 葡萄糖和果糖是叶片中主要的可溶性糖, 而下午13: 00至16: 00,水苏糖成为成熟叶片中含量最高的可溶性糖, 这种变化主要由叶片中水苏糖含量剧烈的昼夜变化引起; 当黄瓜同化物从源叶向外运输时, 韧皮部汁液中的可溶性糖以水苏糖为主, 运输完毕后则以蔗糖为主; 黄瓜成熟叶片在夜间以淀粉、葡萄糖、果糖、水苏糖、蔗糖等各种形式贮藏尚未运出的同化物, 而此时韧皮部汁液中葡萄糖、果糖、水苏糖的含量极低, 可能有特定的源库调节机制阻止叶片中的可溶性糖在夜间进一步向韧皮部中装载。     

弱光胁迫对鲜食甜玉米籽粒糖分积累及产量的影响

为明确弱光对甜玉米品质相关酶活性及产量的影响,以甜玉米品种晶甜9号为试验材料,研究不同遮光率、不同遮光时期对甜玉米籽粒糖分积累、蔗糖合成相关酶及产量的影响。结果表明,随着遮光程度的加深,甜玉米穗位叶蔗糖合成酶和蔗糖磷酸合成酶的活性,花后各阶段籽粒蔗糖、可溶性糖积累量,以及鲜果穗产量均逐渐降低,表现为：遮光率30%>遮光率50%>遮光率80%。在甜玉米生长发育进程中,各时期遮光处理花后穗位叶蔗糖合成酶和蔗糖磷酸合成酶的活性,花后各阶段蔗糖、可溶性糖积累量,以及鲜果穗产量总体表现为：全生育期遮光<大口至吐丝期遮光<吐丝至吐丝后25 d遮光<拔节至大口期遮光<出苗至拔节期遮光。不同遮光率和不同遮光时期共同作用下,全生育期遮光80%处理对玉米籽粒糖分积累及产量的影响最大。综上,不同遮光率、不同遮光时期均显著限制了甜玉米的产量和品质提升,且大口至吐丝期遮光是关键时期。     

黄瓜富氢水浸种对低温下幼苗光合碳同化及氮代谢的影响

为探明外源氢气（H2）对低温下黄瓜幼苗光合碳同化及氮代谢的影响,以‘津优 35 号’黄瓜为试材,将种子分别用饱和富氢水（HRW,H2 供体）和去离子水（对照,Control）浸种 8 h,常温下育苗,幼苗长至 2 叶 1 心时移至光照培养箱中进行低温（昼/夜温度 8 ℃/5 ℃）处理。结果表明：低温抑制黄瓜幼苗的生长,造成叶片光合色素含量、光合速率（Pn）、气孔导度（Gs）、蒸腾速率（Tr）、光下实际光化学效率（ΦPSⅡ）、PSⅡ。最大光化学效率（Fv/Fm）、光下最大天线转换效率（Fv′/Fm′）、核酮糖–1,5–二磷酸羧化酶（RuBPCase）活性和根系活力逐渐降低,而胞间 CO2 浓度（Ci）和初始荧光（F0）趋于升高。此外,低温胁迫诱导了黄瓜幼苗碳代谢关键酶蔗糖合成酶（SS）和蔗糖磷酸合成酶（SPS）活性,总糖、蔗糖含量有所升高,淀粉含量显著下降,同时发现,低温下叶片的硝酸还原酶（NR）、谷氨酰胺合成酶（GS）、谷氨酸合成酶（GOGAT）和谷氨酸脱氢酶（GDH）活性及总氮、铵态氮含量先升高后降低,硝态氮含量略有升高。富氢水浸种的黄瓜幼苗各指标的变化趋势与对照一致,但低温胁迫过程中除 Ci 和 F0显著低于对照外,其余指标多显著高于对照,说明外源 H2 可以通过提高黄瓜幼苗光合关键酶活性,减轻光抑制,维持较高的碳、氮代谢水平,进而增强对低温胁迫的耐性。     

低温弱光下aga2突变体西瓜果实糖代谢差异表达基因分析

为了探明低温弱光环境对西瓜果实糖分积累的调节机制,本试验通过分析棉籽糖水解酶碱性α-半乳糖苷酶基因aga2突变体转录组数据发现低温弱光下aga2突变体西瓜果实中有686个基因上调表达,990个基因下调表达,进一步通过GO和KEGG分析发现MYB、NAC、WRKY等转录因子上调表达抵御低温弱光;而肌醇半乳糖苷合酶(Cla009222)、α-半乳糖苷酶(Cla022883、Cla019238)等与糖分代谢相关的基因在低温弱光下下调表达可能是导致aga2突变体果实糖含量下降的主要原因。本试验通过分析转录组数据,挖掘响应低温弱光的西瓜糖分积累相关基因,为西瓜果实响应低温弱光的糖代谢调控网络奠定了基础。     

冷诱导基因转录因子CBF1转入黄瓜的研究

以抗冷性强的黄瓜"山东5号"为试材,研究冷诱导转录因子CBF1基因对黄瓜抗冷性的影响。从"山东5号"黄瓜基因组DNA中扩增并克隆了冷诱导转录因子CBF1基因,将其与CaMV 35S启动子和Nos终止子融合后构建成植物表达载体pROK2-CBF1。通过花粉管通道法转化黄瓜植株,获得了具有卡那霉素抗性的黄瓜再生植株。结果表明:CBF1基因已整合到黄瓜基因组中,转基因植株胁迫期间可溶性糖含量、幼苗含水量显著高于对照;MDA含量、叶片电解质渗透率显著低于对照,黄瓜已具备了较强抗冷性。     

‘巨峰’葡萄盛花后弱光胁迫对叶片光合生理及光合酶基因表达的影响

为了探明弱光胁迫对葡萄叶片光合生理的影响,以4年生‘巨峰’葡萄为材料,在盛花后进行20 d 70%遮光作为弱光处理,然后恢复至未遮光植株(对照)水平,测定果实品质和叶片的光合参数、叶绿素荧光、叶绿素含量、叶片糖含量及组分、光合酶基因及蔗糖磷酸合成酶基因表达等。结果发现,弱光处理导致成熟期葡萄果实可溶性固形物含量降低,可滴定酸含量升高,果皮花色苷含量减少,果实纵径降低。随着弱光时间延长,叶片叶绿素b含量增加,叶绿素a/b值下降,光合速率和光化学效率降低,叶片中可溶性糖含量降低,Rubisco大、小亚基,PRKase,SPS1-2,SPS3等基因表达受到抑制;光照恢复至对照水平10 d后,光合速率、光化学效率及可溶性糖含量略有上升,光合酶基因表达得到部分恢复;恢复光照50 d后,叶绿素a/b、叶片可溶性糖含量、部分光合酶基因和SPS表达等基本与对照水平一致,仅Rubisco大亚基1(RbcL1)表达仍低于对照。说明长时间弱光处理对葡萄叶片光合中心的损伤短期内不可逆,部分光合酶和蔗糖代谢关键酶基因表达受到抑制,叶片光合能力下降,光合产物合成与积累减少,进而导致了葡萄果实变小,品质降低。     

光质对番茄幼苗碳氮代谢及相关酶活性的影响

采用LED光源,研究了白光（对照）、红光、蓝光、紫光和红蓝（3︰1）组合光对番茄幼苗生长、碳氮代谢及相关酶活性的影响。结果表明：与白光相比,紫光明显抑制幼苗生长,红光抑制幼苗,尤其是其地下部的生长,红蓝组合光下幼苗干物质积累量及壮苗指数最高;在紫光下叶片净光合速率（Pn）、RuBP羧化酶活性、总糖和淀粉含量最低,红蓝组合光下最高;红光和紫光下转化酶的活性较高;红、蓝、紫、红蓝组合光均显著提高蔗糖合成酶（SS）活性,以红蓝组合光更明显,但红、蓝、紫光显著降低蔗糖磷酸合成酶（SPS）活性;红光显著降低了硝酸还原酶（NR）、谷氨酰胺合成酶（GS）和谷氨酸合成酶（GOGAT）活性,蓝、紫、红蓝组合光则显著提高了GS、GOGAT活性以及叶片游离氨基酸和可溶性蛋白含量。总之,红蓝组合光有利于番茄幼苗生长,促进碳同化及总糖和淀粉积累,提高氮代谢相关酶活性及可溶性蛋白和游离氨基酸含量。     

Changes in carbohydrate levels and their metabolic enzymes in leaves,phloem sap and mesocarp during cucumber (Cucumis sativus L.) fruit development

Levels of carbohydrates and activities of metabolic enzymes were examined in leaves (source), phloem sap (flow) and mesocarp tissues (sink) in the course of cucumber (Cucumis sativus L.) fruit development, from 2 days before anthesis to 20 days after anthesis. While total sugar levels increased in all the three sampling organs, starch levels declined in leaves and mesocarp tissues as fruit development progressed. Glucose and fructose were the primary contributors to the soluble sugar pools in mature leaves. Stachyose was found as the most important component of the phloem sap extracts, followed by sucrose and raffinose. However, the primary sugars accumulated in mesocarp tissues were glucose and fructose, not stachyose or sucrose. Activities of sucrose synthesizing enzymes (sucrose phosphate synthase plus sucrose synthase in the synthesizing direction) exceeded that of sucrose degrading enzymes (acid invertase, neutral invertase plus sucrose synthase in the degrading direction) in leaves, which might cause a sucrose pool utilized in raffinose and stachyose biosynthesis. While alkaline a-galactosidase form I activity declined, stachyose synthase activity showed a rapid increase until 12 days after anthesis and only subsequently decreased in leaves. Activities of sucrose degrading enzymes were always much higher than that of sucrose synthesizing enzymes in mesocarp tissues. Thus, sucrose accumulation could not occur in mesocarp tissues. While stachyose synthase activity steadily decreased, alkaline a-galactosidase form I activity showed a moderate increase before decrease in mesocarp tissues. The relationship between levels of soluble sugars and activities of relative enzymes was also discussed.     

Low night-temperatures affect the metabolism of raffinose-family oligosaccharides in melon (Cucumis melo L.) leaves during fruit expansion

Summary

This study served to reveal the effects and mechanism of low night-temperatures on the metabolism of raffinose-family oligosaccharides in melon (Cucumis melo L.) leaves. We investigated the carbohydrate contents and sugar-metabolising enzyme activities in leaves under normal (15°C; control) and two low night-temperature (12°C or 9°C) conditions during fruit expansion. The results showed that the greatest difference in carbohydrate metabolism occurred between 9°C at night and the 15°C control. Compared to the normal night temperature, total carbohydrate accumulation, in particular raffinose, increased and the ratio of raffinose to total carbohydrates exhibited the greatest increase in melon leaves treated at 9°C at night, while the ratio of sucrose to total carbohydrates declined at the end of the same treatment. These data suggested that 9°C at night played an important role in partitioning carbon from sucrose to raffinose-family oligosaccharides (RFO), and that raffinose may serve dual purposes in stress protection and carbon storage. Meanwhile, the activities of sugar-metabolising enzymes were notably affected by 9°C at night. Sucrose synthase (SS), sucrose phosphate synthase (SPS) and, in particular, galactinol and stachyose synthase activities decreased markedly, while acid invertase (AI) and neutral invertase (NI) activities increased sharply, which restricted sucrose and RFO synthesis. Overall, these data suggest that the decline in stachyose and galactinol synthase activities were the main limiting factor for RFO synthesis in melon leaves at 9°C at night. They also indicated that the reduction in stachyose metabolism may be an important factor during the development of unmarketable fruit.     

Salicylic acid regulates sugar metabolism that confers tolerance to salinity stress in cucumber seedlings

Salicylic acid (SA) plays a critical role in plant development and defense responses to biotic and abiotic stresses. Sugars can act as osmotic adjustors or nutrient and metabolic signals in the activation of plant defense responses. To uncover the effects of SA on sugar metabolism, we assayed the altered components and levels of sugars in cucumber seedlings treated with SA. After SA treatment, the levels of glucose, fructose, raffinose and stachyose were increased in both leaves and roots. In contrast, sucrose and starch responded differently in leaves and root, decreasing in leaves but accumulating in roots. These changes could be due to the effects of SA on the activities of metabolism-related enzymes. In leaves, SA promoted the activities of sucrose phosphate synthase (SPS), sucrose synthase (SS) and amylases, while the SA-treated root showed a reduced amylase activity and an unchanged activity in SPS. The changes in various sugar contents resulted in the accumulation of soluble sugars in SA-treated cucumber seedlings, especially non-reducing sugars in roots. These increased sugars could function as osmotic regulators and facilitate water uptake and retention in plant cells, thereby conferring seedlings an enhanced tolerance to salinity stresses caused by NaCl treatment. Taken together, our findings provide an important insight into the effects of SA on sugar metabolism, and a protective mechanism for SA against water deficiency is discussed.     

低氮胁迫下黄瓜钙依赖蛋白激酶基因的克隆及表达分析

在低氮胁迫下，以黄瓜品种津研4 号叶片为供试材料，以cDNA 为模板，依据黄瓜基因组数
据库中Csa002986 基因编码区全序列，应用Primer Premier 5.0 软件设计引物，克隆得到黄瓜钙依赖蛋白
激酶基因（calcium-dependent protein kinase，CDPK）。该基因全长1 584 bp，编码527 个氨基酸。预测
该基因编码的蛋白是一个稳定的亲水蛋白，无跨膜结构，无信号肽，存在蛋白激酶结合区、EF 手型钙结
合区、丝氨酸/苏氨酸蛋白激酶活性位点、N 酰基化位点等多个位点。CDPK 基因在不同氮浓度处理下黄瓜
各部位表达分析结果显示，在无氮条件下，CDPK 基因在茎尖表达量最高，其次为叶和茎；在低氮条件下，
CDPK 基因在茎中表达量最高，其次为茎尖和叶；在正常及高氮条件下，CDPK 基因在茎尖表达量最高，
其次为茎和叶。CDPK 基因在叶中的表达模式分析结果显示，在无氮及低氮胁迫下CDPK 基因表达量增加，
随着氮浓度的降低，CDPK 基因的表达量增加并明显高于对照；在高氮胁迫条件下，CDPK 基因的表达量
低于对照。     

Different mechanisms to obtain higher fruit growth rate in two cold-tolerant cucumber (Cucumis sativus L.) lines under low night temperature

One cold-sensitive cultivar (Jinyan 4) and two cold-tolerant inbred lines (NY-1 and XC-1) of cucumber (Cucumis sativus L.) were subjected to temperatures of 28 °C/22 °C (day/night, control) or 28 °C/12 °C (day/night, cold treatment) in a 10 h photoperiod (7:00–17:00). Under control conditions, cucumber fruits grew fast during afternoon and early night, and slow during late night and morning. Under 28 °C/12 °C conditions, the two cold-tolerant inbred lines maintained relatively higher fruit growth rates than the cold-sensitive cultivar by different mechanisms. Compared to Jinyan 4, NY-1 fruits had higher growth rates during cold nights while XC-1 fruits grew faster during the next day. Under the 28 °C/12 °C temperature regime, the assimilate accumulation in the fruits of all tested genotypes followed a similar trend with the corresponding fruit growth rates. After a cold night treatment, the net CO₂ assimilation rates of one- and two-fruit plants, which had increased sink demand, were higher than that of plants without fruits in all tested genotypes. This response indicates that feedback inhibition might be an important reason for the reduction of photosynthesis on the next day. In addition, after a cold night treatment, the levels of exportable sugars (sucrose and stachyose) in mature leaves of XC-1 were higher than those measured in Jinyan 4 and NY-1, which might explain why XC-1 fruits had faster assimilate accumulation rates in the next morning. Higher activity of sucrose-phosphate synthase, a key enzyme of sucrose and stachyose biosynthesis, constituted an additional evidence that faster sucrose and stachyose biosynthesis in mature leaves may occur in XC-1 than in Jinyan 4 and NY-1 at that time. In conclusion, our results showed that cucumber genotypes may use different mechanisms to enhance their cold tolerance.     

黄瓜细胞分裂素合成关键酶IPT 基因家族序列特征及其表达分析

 为了分析黄瓜IPT家族基因,以及黄瓜IPT基因CsIPT与果实发育的关系,以黄瓜单性结实自交系‘EC1’和非单性结实自交系‘8419s-1’为试材,应用荧光定量PCR技术,在分析不同器官中CsIPT表达特征的基础上,寻找果实发育中的作用基因,进一步分析这些基因在果实发育过程中的表达特征。结果表明：8个黄瓜IPT基因（CsIPT1 ~ CsIPT8）核苷酸相似性在17.3% ~ 33.2%范围内,氨基酸长度在280 ~ 504 AA之间,具有共同的ATP/GTP结合功能域[(A,G)-X₄-G-K-(S,T)],CsIPT4氨基酸序列中包含有一个锌指基序（C-X₂-C-X_12,18-H-X₅-H）类似序列;黄瓜幼果中CsIPT3、CsIPT5、CsIPT7和CsIPT8表达水平较高;CsIPT1、CsIPT3、CsIPT4和CsIPT5的表达量在未发育的果实中有增加的趋势,CsIPT2和CsIPT8在未发育的果实中表达基本无变化,而在授粉后发育的果实中表达量很高,说明这两个基因可能与授粉后黄瓜果实发育调控有关。此外,CsIPT2在天然单性结实材料‘EC1’果实发育早期表达较高,推测其在黄瓜单性结实果实发育中发挥着重要作用。     

黄瓜根结RDR酶基因的分离与表达分析

 基于黄瓜（Cucumis sativus L.）自交系9930全基因组序列,采用RT-PCR的方法,分离出根结形成过程中表达的RDR基因,共获得了5个CsRDR基因：CsRDR1a、CsRDR1b、CsRDR1c、CsRDR2和CsRDR6（HQ738485 ~ HQ738489）。qPCR定量分析表明,5个基因在接种根结线虫后的不同时期表达量都有不同程度的增加。CsRDR1a、CsRDR1b、CsRDR2和CsRDR6在接种线虫早期（6 h）相对表达量明显增加;而CsRDR1c在接种36 h时相对表达量达到最高,从而表明CsRDR基因与黄瓜根结的发育相关。