棉花中GhTFL1a和GhTFL1c基因的表达及启动子分析

从陆地棉中克隆了磷脂酰乙醇胺结合蛋白GhTFL1a和GhTFL1c基因,并对该基因进行表达分析、启动子预测和启动子活性研究。利用启动子分析软件PlantCARE预测得出,GhTFL1a启动子区域有脱落酸响应元件、干旱诱导的MYB结合位点和顶芽特异表达响应元件等;GhTFL1c启动子区域有乙烯响应元件、干旱诱导的MYB结合位点和水杨酸响应元件。因此,将pGhTFL1a和pGhTFL1c分别构建到启动子检测载体pBI121-GUS上形成融合表达载体,通过烟草瞬时转化检测得出这2个基因的启动子都具有活性。实时荧光定量PCR分析表明, GhTFL1a和GhTFL1c在光周期处理和不同材料的陆地棉(栽培种和半野生种)中表达模式呈相反趋势。GhTFL1a基因受脱落酸(abscisic acid, ABA)、水杨酸(salicylic acid, SA)和盐胁迫诱导,而GhTFL1c可以响应赤霉素(gibberellin, GA)、SA和ABA胁迫。研究结果初步表明,GhTFL1a和GhTFL1c可能参与了植物逆境胁迫脱落酸和水杨酸响应的调控,为在棉花中进一步阐明其功能奠定了基础。     

不同番茄材料黄化曲叶病毒病抗性评价及抗性基因检测

以15份自育的无限生长型番茄材料为试材，抗番茄黄化曲叶病毒（Tomato yellow leaf curl virus,TYLCV）品种‘齐达利’和感病品种‘Money Maker’(MM)为对照，采用苗期农杆菌接种法，研究接种黄化曲叶病毒后的病情指数，幼苗叶片防御酶活性的变化，不同基因型番茄材料的抗性水平。结果表明：853、857、867表现为免疫；817、842表现为高抗；805、818、819、820、841表现为中抗；801、802、803、822为感病。853、857、867的防御酶(PAL、PPO、POD)活性在发病初期均达到峰值，且整体高于其他接种材料，其他接种材料酶活性峰值则出现较晚。含抗性基因的接种材料均表现出不同程度的抗性，含有纯合 Ty-1和 Ty-3基因的853、857、867对 TYLCV抗性更高。经综合评价，853、857、867可作为抗番茄黄化曲叶病毒病的优良番茄材料，817、842可作为备选材料。     

生防细菌复配对苹果主要病原真菌抑菌活性筛选及其稳定性

采用生长速率法测定4种生防细菌及其复配对5种苹果主要病原菌的抑制效果，并将筛选出的最佳复配生防细菌发酵液进行稳定性测定。结果表明，单一生防细菌发酵液中多粘类芽孢杆菌FS-1206对苹果树腐烂病菌的抑菌效果最好，为78.19%；当多粘类芽孢杆菌FS-1206与解淀粉芽孢杆菌TS-1203复配比例为 2∶1时，其对苹果树腐烂病菌的抑菌率高达86.64%，抑菌效果显著优于单一生防细菌发酵液。稳定性测定结果表明，复配生防细菌（FS-1206∶TS-1203= 2∶1）发酵液在紫外照射、高温和强酸强碱环境条件下，其抑菌活性均具有良好的稳定性。     

基于叶片质外体相关成分和酶活性响应的龙葵耐Cd机理研究

为探讨龙葵(Solanum nigrum L.)耐镉(Cd)机理,采用基质培养法,研究了50 μmol·L^-1 Cd(CdCl₂·2.5H₂O)处理120 d对其生长和叶片细胞壁果胶、木质素含量、多聚半乳糖醛酸酶(PG)、纤维素酶(Cx)、NADH-过氧化物酶(NADH-POD)、愈创木酚过氧化物酶(GPOD)和松柏醇过氧化物酶(CAPX)活性以及质外体汁液中过氧化氢(H₂O₂)含量和GPOD、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和超氧化物歧化酶(SOD)活性的影响。结果表明,与对照相比,Cd处理显著抑制了龙葵地上和地下部的伸长生长、降低了根的干物质积累(P<0.05),但茎的分枝增多,叶面积、叶绿素含量、净光合速率和胞间CO₂浓度无显著变化(P>0.05);细胞壁结合酶NADH-POD、GPOD和CAPX活性以及果胶含量显著增高(P<0.05),而PG活性显著下降(P<0.05),Cx活性和木质素含量无显著变化(P>0.05);质外体汁液中H₂O₂含量显著增加(P<0.05),GPOD和SOD同工酶谱及活性无显著变化(P>0.05);CAT同工酶谱带增宽,APX同工酶谱带数增加,二者活性均显著升高(P<0.05)。说明50 μmol·L^-1 Cd较长时间处理龙葵植株,其地上部分生长仍处于良好状态,表现出对Cd毒害的耐性,这与Cd处理后叶片细胞壁果胶含量和结合酶GPOD以及质外体汁液中APX、CAT活性升高有关,推测这是龙葵耐Cd的一种机理。     

龙须藤茎皮甲醇提取物生物活性初探

龙须藤Bauhinia championii在医用方面已证实具有良好的抗菌、消炎作用,但其在农用方面的活性研究还鲜有报道。本研究分别采用菌丝生长速率法和种子生物测定法,首次测定龙须藤茎皮甲醇提取物及分离组分对6种农业有害真菌和单、双子叶两种植物的生物活性。结果表明,龙须藤甲醇浸提物对水稻稻瘟病菌Magnaporthe oryzae有一定的抑制效果,对油菜Brassica napus和稗草Echinochloa crusgalli的根芽均没有抑制作用。但其甲醇浸提物中分离的大部分组分对供试病原菌和植物的根芽都表现出一定抑制作用。杀菌活性方面:10号组分对西瓜枯萎病菌Fusarium oxysporum和番茄早疫病菌Alternaria solani的抑制效果最好,抑制率分别达到80.0%和78.0%,除草活性方面:1～3号组分对油菜根的抑制率较高,达到50%以上,2号组分对稗草根的抑制效果最好,达到68%。可见,龙须藤茎皮提取物中含有具有良好杀菌活性和除草活性的物质。     

链霉菌和壳聚糖对淮山土壤微生物活性与群落的影响

为探究链霉菌30702和壳聚糖对淮山土壤微生物活性与群落的影响,采用比色法和滴定法观测土壤酶活性的变化,采用高通量测序技术和Trimmomatic等软件,分析土壤微生物群落中细菌和真菌的发展变化。结果表明,链霉菌、壳聚糖、土壤原始微生物和培养时间等4个因素对淮山土壤的脲酶和过氧化氢酶活性均有显著影响,壳聚糖浓度在0~10.0 g/kg内,土壤酶活性随壳聚糖浓度的增大而提高;随时间的延长,先升再降;壳聚糖与链霉菌之间存在交互效应。链霉菌和壳聚糖能增加淮山土壤细菌的物种丰度,减少细菌的多样性,增加艾德昂菌属（Ideonella）和纤维弧菌（Cellvibrio）等有益菌属的相对丰度,而对真菌的物种丰度和多样性的影响较小,与对照之间的差异不显著,土壤优势菌属为腐质霉属（Humicola）、枝孢属（Cladosporium）、鞘氨醇单胞菌属（Sphingomonas）和uncultured_bacterium_ c_Subgroup_6。其中,链霉菌与2.5 g/kg壳聚糖复合处理的细菌物种丰度最大,细菌多样性最少。     

Effect of Foliar GA3 and ABA Applications on Reactive Oxygen Metabolism in Black Currants (Ribes nigrum L.) During Bud Para-dormancy and Secondary Bud Burst

The secondary bud burst can cause around 10％-20％ yield losses in black currants, an economically important crop in parts of Europe, Asia and North America. The metabolism of reactive oxygen species (ROS) has been linked to bud dormancy and its early release (secondary bud burst) in several fruit crops. But the relationship between ROS metabolism and the secondary bud burst is still not well understood in black currants. In the present study, two black currant cultivars (Adelinia and Heifeng) with opposing tendency of exhibiting the secondary bud burst were sprayed with abscisic acid (ABA) and gibberellic acid (GA3) to either inhibit or induce the secondary bud burst. The results showed that ABA inhibited the secondary bud burst by reducing the contents of ROS (H2O2, O2-?) in buds; decreasing the activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT); and increasing the contents of oxidized glutathione (GSSG) and ascorbic acid (AsA). GA3 effectively induced the secondary bud burst by increasing ROS contents; increasing the activities of several antioxidant enzymes, such as SOD, POD, CAT, glutathione reductase (GR), ascorbate peroxidase (APX) and the contents of reduced glutathione (GSH); and decreasing the contents of AsA. The experimental results showed that GA3 treatment increased the content of ROS, accelerated the metabolism of reactive oxygen species, and promoted the second burst of black currants. However, ROS metabolism was at a low level under ABA treatment, and the buds remained dormant. These results suggested that ROS metabolism might play an important role in the two black currants of the secondary bud burst.     

Characterization of Spray-Dried Hydrolyzed Proteins from Pink Perch Meat Added with Maltodextrin and Gum Arabic

In the present study, spray-dried pink perch meat protein hydrolysate (PPH) and hydrolysate with maltodextrin and gum arabic (PPHMG) were characterized in relation to their physical, rheological, functional, antioxidant, thermal, and sensory properties. The Herschel–Bulkley model was suitable to explain the flow behavior of PPH and PPHMG emulsions. Both the samples exhibited functional and antioxidant properties. Antioxidant activities were significantly higher for PPH compared with PPHMG (p<0.05). PPH and PPHMG alone did not exhibit antimicrobial activity but enhanced the activity of chitosan against pathogens. Sensory acceptability of the samples in soup revealed that PPHMG of up to 4% was highly scored without any traces of bitterness, whereas for PPH, a concentration of up to 3% was acceptable.     

橄榄多酚氧化酶基因(PPO)克隆及其试管苗离体保存过程中的表达分析

以来源于橄榄成熟胚的橄榄试管苗为材料,进行PPO基因克隆,比较试管苗不同保存阶段PPO基因转录水平和PPO酶活性的变化。结果表明:克隆获得1个PPO基因全长cDNA序列,命名为PPO2,GenBank登录号为JQ319005,cDNA全长序列为1 934 bp,推测出橄榄PPO开放阅读框(ORF)1 818 bp,编码606个氨基酸。橄榄PPO2属于碱性、亲水、非分泌蛋白,可能定位于线粒体基质空间,具有3个典型的保守结构域,丝氨酸磷酸化占主导地位,并且二级结构以不规则卷曲为主。而三级结构预测表明,橄榄试管苗PPO2蛋白主要由α螺旋和不规则卷曲组成;进化树分析显示橄榄PPO蛋白与胡杨PPO关系最近,而与葡萄和莲的亲缘关系较远;qPCR分析显示,PPO2基因在橄榄试管苗不同保存阶段都有表达,但在橄榄试管苗保存1个月幼叶比较多时PPO2基因的表达量最高;而橄榄试管苗在分别保存1、2、3、4、5个月时PPO活性变化趋势不明显,当保存时间到6、7、8个月时PPO活性急剧增加。因此,PPO基因可能与幼嫩组织的启动分化有关,并在橄榄试管苗生长发育过程中发挥重要作用。