菌根真菌液泡的分离及特性研究

从菌根真菌黑核菌（ＣｅｎｏｃｏｃｃｕｍｇｅｏｐｈｉｌｕｍＦｒ．）和鹅膏菌［Ａｍａｎｉｔａｍｕｓｃａｒｉａ（Ｌ．ｅｘＦｒ．）Ｐｅｒｓ．ｅｘＨｏｏｋ．］菌丝中，采用多元碱化合物诱导原生质体裂解方法，分离出细胞器──液泡。实验结果表明，在加磷的ＭＭＮＣ培养基中培养４—７天的菌丝经水解酶作用所释放出的原生质体具有较高的液泡化程度；液泡产率约为原生质体的５－１０％；细胞质特征酶葡萄糖－６－磷酸－脱氢酶活性测定结果证明了液泡的纯度；而等量原生质体和液泡中酸性磷酸酶及α－甘露糖苷酶活性的比较表明这两种酶主要存在于液泡中，因此，它们可以作为菌根真菌黑核菌和鹅膏菌液泡的特征酶。     

柑橘幼果发育期碳水化合物代谢及其与生长发育的关系

温州蜜柑幼果在盛花后逐渐生长 盛花后6 d开始大量脱落 日本甜夏橙幼果在盛花后12 d内重量未增加，一直处于急剧落果期。甜夏橙幼果中蔗糖含量高出温州蜜柑20多倍，种间还原糖含量变化趋势略有不同。两种柑橘的淀粉含量变化也不同，但它们的淀粉含量与淀粉酶活性、幼果相对脱落率均呈负相关。对幼果碳水化合物代谢与生长、脱落的关系作了讨论。     

银杏幼苗叶片转化酶活性及其动力学

采用波钦诺克XH法研究了田间和室内栽培的银杏幼苗叶片转化酶的活性及其动力学性质。结果表明:银杏幼苗叶片转化酶在pH值5.0、温度55℃时活性最强;该酶的米氏常数为3.7×10-2mol·L-1;室内盆栽的银杏幼苗叶片转化酶的活性显著高于田间栽培的同龄银杏幼苗叶片转化酶的活性。     

Effects of late planting on fiber quality and within-boll yield components as mediated by sucrose metabolism in cotton bolls

ABSTRACT

The objective of this study was to examine the effect of late planting on sucrose metabolism in cotton bolls and the relationship to fiber properties and within-boll yield components. Two cotton lines A201 and A705 were employed in a sowing date experiment where two temperature regimes during boll maturation period were created at mean daily temperature of 26.8°C, 28.3°C for early planting and 25.2°C, 23.1°C for late planting in 2016 and 2017, respectively. Boll size, seed mass per boll, seed index and fiber length were increased, and lint percentage was decreased by late planting. Greater cell wall invertase activity and the resultant hexose concentration in fibers were observed in late planting, and thus led to decreased osmotic potential accounting for the enhanced fiber length. Similarly, late planting increased the maximum of vacuolar invertase activity in ovules occurring at 5 days post anthesis (DPA) and hexose concentrations in embryos from 10 DPA afterwards which may favor embryo cell division, and thus increase final seed size. Our data indicate that acid invertase and hexose are implicated in the formation of within-boll yield components and fiber properties as affected by the lower temperature regime due to late planting.     

Comparison of Salt and Drought‐Stress Effects on Maize Growth and Yield Formation with Regard to Acid Invertase Activity in the Kernels

The long‐term effects of salt stress (11 dS m^?1) and drought stress (35 % WHC) were investigated for two maize genotypes, focusing on the relation between metabolic changes around the time of pollination and the impact on yield determinants at maturity. The relatively salt‐resistant hybrid Pioneer 3906 and the relatively drought‐resistant hybrid Fabregas were compared. The experiments were conducted in large plastic containers in a vegetation hall in two consecutive years (2011 and 2012). Plant height and leaf area were significantly reduced under both stress conditions. The transpiration rate was only slightly reduced under drought stress; but under salt stress, a significant reduction occurred 40–53 days after sowing. As a significant increase in sucrose concentrations was observed in the salt‐treated maize kernels 2 days after pollination, the availability of assimilates was not limiting and the plants could afford to save water by reduced stomatal opening. Although under both stress conditions the soluble acid invertase activity was reduced 2 days after pollination, concomitantly, an increase in hexose concentrations was observed. Thus, in these experiments, the delivery of hexoses by acid invertase activity did not limit kernel development. Differences in grain yield at maturity between salt and drought stress were most likely caused by salt‐specific effects (Na⁺ toxicity), Fabregas being more affected than Pioneer 3906.     

菠萝酸性转化酶基因家族初步生物信息学及表达分析

酸性转化酶（acid invertase, AIN）在菠萝采后蔗糖降解过程中起着重要作用,基于菠萝全基因组数据库,预测菠萝AIN家族基因并进行生物信息学分析,解析其在采后菠萝不同贮藏温度下的表达变化情况,为阐明AIN基因在采后菠萝果实贮藏特性中的作用奠定基础。以水稻AIN家族基因为探针,在菠萝全基因组中鉴定到2个菠萝细胞壁酸性转化酶基因（cell wall acid invertase, CWIN）和2个液泡酸性转化酶基因（vacuolar acid invertase, VIN）,分别命名为AcCWIN1、AcCWIN2、AcVIN1、AcVIN2,设计编码区引物进行测序验证,并进行生物信息学分析。进化分析结果表明,AcCWIN1、AcCWIN2和AcVIN1、AcVIN2蛋白分别归于细胞壁酸性转化酶和液泡酸性转化酶2个进化支上,且均属于糖基水解酶家族GH32,基因结构、保守域和保守基序均一致。荧光定量分析结果表明,菠萝果肉中AcVIN1和AcVIN2在果实采后贮藏过程中表达量升高,且AcVIN1在发生黑心病的部位大量表达,而AcCWIN1和AcCWIN2在采后贮藏过程中表达量逐渐降低,且随着贮藏温度的升高其表达量降低,预示AcVIN1、AcVIN2较AcCWIN1、AcCWIN2在菠萝采后蔗糖降解和黑心病的发生方面发挥着更为重要的作用。     

蔗糖转化酶家族基因进化、表达及对草莓果实糖分积累的影响

在草莓蔗糖碱性/中性转化酶家族基因(Fa.A/N-Inv)成员序列分析的基础上,以‘申阳’和‘红颜’草莓为材料,利用实时定量PCR技术分析了Fa.A/N-Inv家族基因在不同条件下的表达模式,利用分光光度法测定了果实发育过程中A/N-Inv酶活性,利用高效液相色谱法分析了果实发育阶段糖分积累规律。结果表明:(1)Fa.A/N-Inv是由8个成员组成的多基因家族;(2)4个Fa.A/N-Invs不仅具有组织特异性,还具有果实发育阶段特异性和品种特异性表达特点;(3)4个Fa.A/N-Invs的表达受细胞分裂素(BA)、赤霉素(GA)和生长素(IAA)诱导;(4)蔗糖和葡萄糖作为A/N-Inv酶促反应的底物和产物,分别对4个Fa.A/N-Invs转录表达起正、负反馈调节作用,此外蔗糖还可作为一种信号分子诱导Fa.A/N-Invs表达;(5)‘申阳’草莓A/N-Inv酶活性在绿果阶段低于‘红颜’,其余阶段均高于‘红颜’;(6)成熟果实中蔗糖含量‘红颜’(50.7 mg·g~(-1) FW)约为‘申阳’(23.2 mg·g~(-1) FW)的2倍。     

尕海湿地区沼泽草甸退化对土壤氮转化酶活性的影响

为探讨高寒湿地退化对土壤氮转化酶活性的影响,以青藏高原东缘尕海湿地未退化（ND）、轻度退化（LD）、中度退化（MD）和重度退化（HD）4种不同退化程度0~40 cm土层沼泽草甸为研究对象,研究不同退化与土层中土壤氮转化酶（蛋白酶、脲酶、硝酸还原酶和亚硝酸还原酶）活性的变化特征及其与土壤理化性质之间的关系。结果表明：1）随沼泽草甸退化程度加剧,土壤含水量、全氮、铵态氮和微生物生物量氮含量均显著降低,土壤温度与硝态氮含量却显著增加。2）随退化程度加剧,各土层土壤脲酶活性增加、蛋白酶活性降低,且仅在20~40 cm土层存在显著差异;硝酸还原酶活性增加、亚硝酸还原酶活性降低,在0~20 cm土层存在显著差异。3）各退化程度中,土壤脲酶、蛋白酶、亚硝酸还原酶活性均随土层深度的增加而显著下降,硝酸还原酶活性仅在HD显著下降。4）退化程度和土层对4种土壤氮转化酶活性均存在显著影响,且对土壤硝酸酶和亚硝酸还原酶活性存在显著交互作用。5）冗余分析表明土壤含水量对土壤氮转化酶活性变化的贡献率高达67.1%,其是驱动尕海沼泽草甸退化演替过程中土壤氮转化酶活性变化的主导因素。研究结果可为高寒湿地生态系统退化中的土壤酶活性变化规律提供理论依据。     

杨梅酸性转化酶基因cDNA分离及表达分析

杨梅果实富含蔗糖,酸性转化酶是蔗糖代谢关键酶,根据植物酸性转化酶基因保守区序列设计引物,提取杨梅叶片RNA,逆转录获得cDNA,以此为模板通过PCR技术扩增到长度为516bp的基因片段,克隆入pMD18-T载体中,命名为MrIVR1(GenBank:DQ339699)。测序及同源性检索表明,该基因推导氨基酸序列与君子兰、葡萄、草莓、胡萝卜等酸性转化酶基因氨基酸序列同源性为60%～69%。运用ClustalX软件对植物转化酶基因进行了系统树分析,结果显示,MrIVR1编码的蛋白质属于细胞壁酸性转化酶。半定量RT-PCR表达分析显示,MrIVR1基因在杨梅果实发育早期表达量最高,随着果实的发育表达量下降,在成熟果实中表达水平较低。     

赤霉素对发育的大豆种子中同化物卸出和积累的影响

用微注射法将不同浓度的赤霉素 (GA3 )引入开花后 30 d的大豆种子幼胚的 2片子叶中间的质外体空间 ,同时给叶片饲喂 14 CO2 ,以种皮和子叶中放射性强度来比较分析 GA3 对同化物卸出的影响。分析了种皮和子叶中的酸性转化酶活性 ,发现 GA3 1× 10 -5mol.L-1使种皮中酸性转化酶活性提高而子叶中酶活性降低 ,从而促进种皮同化物的卸出和向子叶的分配。实验结果还表明 ,引入 GA3 影响了种子内源脱落酸 (ABA)水平 ,而且子叶中的 ABA含量变化和同化物积累相关 ,GA3 1× 10 -5mol.L-1使子叶中 ABA水平上升 ,伴随总糖、蔗糖和蛋白质的积累增加 ,GA3 1× 10 -4 mol.L-1却使 ABA水平下降 ,而且物质积累也减少