褪黑素诱导小豆抗锈病机理的初步研究

为明确外源褪黑素诱导小豆抗锈性的作用及机理,以感病小豆品种‘宝清红’为材料,采用叶面喷施不同浓度褪黑素激发处理小豆真叶,而后对真叶挑战接种锈菌夏孢子,结果表明,低浓度(11.61 mg/L)褪黑素可显著提升小豆对锈病的抗性。夏孢子萌发试验表明,褪黑素对夏孢子萌发及芽管生长无显著抑制作用,表明褪黑素无抑菌活性。进一步的基因表达分析发现,与对照相比,褪黑素激发诱导了水杨酸(SA)通路关键基因NPR1于接种后24 h显著上调表达,且病程相关蛋白PR1、几丁质酶(CHI)、β-1,3-葡聚糖酶(GLU)及PR5均于接种后24～120 h被显著诱导表达,说明褪黑素可能通过诱导NPR1表达,进而激活下游PR蛋白的高水平应答,使感锈病小豆品种获得对锈病的抗性。     

Grain Quality as Affected by Down-regulation of Expression of Different ALK Alleles in indica Rice (Oryza sativa L.)

【Objective】Gelatinization temperature (GT) is one of the key physicochemical properties in rice quality, which is mainly regulated by ALK (SSII-3) gene. In general, there are two ALK alleles among indica cultivars. To detect their functional differentiation in indica rice,【Method】Zhenshan 97B (a high GT variety carrying ALKc allele) and Longtefu B (a low GT variety carrying ALKb allele), were used as receptors for the generation of transgenic rice with down-regulation of ALK expression by RNA interference (RNAi).【Result】Down-regulation of ALK gene significantly decreased the GT of the transgenic lines. Due to the difference of original GT between the two receptors, the GT of transgenic rice lines derived from Zhenshan 97B (a high GT variety) decreased significantly, but it is slightly decreased in transgenic plants derived from Longtefu B (a low GT variety). The differential scanning calorimetry (DSC) results showed that the initial temperature of RNAi transgenic rice was significantly lower than the corresponding control and the transgenic lines were gelatinized in advance. The peak value of GT(Tp) in RNAi rice grains was significantly lower than that of the control under Zhenshan 97B background. However, Tp of RNAi rice grains under Longtefu B background was significantly lower than the control to a lesser extent. Also, down-regulation of ALK expression had a significant effect on rice physical-chemical characteristics. An increase of apparent amylose content in RNAi transgenic plants was detected due to the decreased expression of ALK gene. Besides, the pasting properties showed that down-regulation of ALK gene had obvious effects on peak viscosity and breakdown value, improving the taste of the transgenic rice. The gel consistency was significantly different among Zhenshan 97B RNAi lines and their parents, but no difference was found in Longtefu-derived transgenic lines.【Conclusion】RNA interference to ALK allele expression had a significant effect on rice quality, especially the gelatinization characters. Down-regulated expression level of ALKc allele would cause larger variation of physical-chemical characteristics between transgenic rice and their parent than that of ALKb allele.     

36个小麦审定和区试品种(系)的抗白粉病基因推导

明确我国当前小麦审定和区试品种含有的抗白粉病基因, 可为这些品种在小麦抗病育种中的应用及品种合理布局和轮换提供依据。本研究采用24个不同毒性的小麦白粉菌菌株对36个小麦审定和区试品种(系)进行抗白粉病基因推导, 参试品种(系)与46个已知抗病基因小麦品种(系)抗谱比较的结果表明, 11个小麦审定品种中有5个品种对所有供试白粉菌菌株表现抗性, 结合亲本溯源, 推测其中有3个品种可能携有Pm21基因; 另外6个审定品种中有5个品种可能含有抗病基因Pm2, 且其对应的亲本或亲本组合中含有抗病基因Pm2。25个区试品种(系)中有3个可能含有Pm21, 10个含有Pm2, 1个含有Pm2+6,2个含有Pm4b,1个含有Pm8。另外, 参试的36个品种(系)中还有9个品种(系)和已知基因品种抗谱存在一定差异。总体上, 推导出已知基因的品种以含有Pm2基因的品种最多, Pm21基因的品种次之, 建议在生产上加强对Pm21基因品种(系)特别是已审定的携有Pm21基因品种的推广和应用, 应该注意一些省份在育种和生产上应慎用或少用含Pm2基因的品种(系)。     

粳稻柱头外露率QTL定位

【目的】柱头外露是影响杂交水稻制种的一个重要性状。本研究旨在初步鉴定控制柱头外露率的QTL。【方法】以高柱头外露率粳稻两系不育系大S×低柱头外露率热带粳稻品系D50的F_2群体及海南陵水、浙江杭州两地重复种植的F_(2:3)群体为材料,考查F_2群体及陵水、杭州两地两次重复F_(2:3)群体单边柱头外露率(PSES)、双边柱头外露率(PDES)和柱头总外露率(PES)。【结果】相关分析表明,单边柱头外露率、双边柱头外露率和柱头总外露率之间存在极显著的正相关。根据构建的大S×D50 F_2群体遗传连锁图谱,QTL分析检测到水稻柱头外露率QTL 21个,分布于第1、2、3、4、6、7和12染色体上,贡献率变幅为0.1%~57.6%,除个别贡献率极低的位点外,绝大部分QTL加性效应来自于高值亲本大S。其中位于第4染色体和第12染色体的两个位点q PDES4,q PES12在F_2和两次重复F_(2:3)群体中均检测到,贡献率分别达17.86%和16.98%。位于第3染色体的q PES3位点在陵水、杭州两次F_(2:3)群体中重复检测到,贡献率高达25.6%。【结论】检测到的QTL,有些与其他学者报道的处于同一位置,有些则是新的柱头外露率QTL。这些水稻柱头外露率QTL为今后柱头外露率基因精细定位、克隆、以及利用分子标记辅助选择(MAS)选育高柱头外露率粳型不育系提供理论依据。     

夜蛾黑卵蜂雌雄蜂鉴别方法及雌蜂寄生草地贪夜蛾卵的行为特征

夜蛾黑卵蜂Telenomus remus被认为是草地贪夜蛾Spodoptera frugiperda卵期的高效寄生蜂。明确夜蛾黑卵蜂基本生物学特征将为其繁育及应用提供重要参考。本研究对雌、雄蜂外部形态及雌蜂寄生行为进行了观察。结果显示,1、除雌蜂产卵器和雄蜂外生殖器等第一性征外,夜蛾黑卵蜂雌、雄蜂个体还具备一些典型的第二性征,可通过触角环节数量、触角棒节和柄节形态以及足和触角的颜色进行区分。2、在同一卵块上,雄蜂先于雌蜂羽化,并撕咬寄主卵壳协助雌蜂羽化;雌蜂羽化后雄蜂会立即求偶并与其交配。3、夜蛾黑卵蜂雌蜂通过利用寄主卵表鳞毛空隙、主动清理鳞毛和移动表层卵粒3种行为策略实现对大部分草地贪夜蛾卵的寄生。本研究建立了夜蛾黑卵蜂雌、雄蜂的鉴别方法,并明确了雌蜂应对夜蛾黑卵蜂卵表结构防御的行为策略。     

Effects of Exogenous Materials on Nitrogen Photosynthetic Efficiency and Yield Formation of japonica Rice in Cold Region Under Cold Water Stress at Booting Stage

【Objective】Our goal is to develop disaster-reducing, production-preserving and chemical-controlling cultivation techniques to effectively resist cold damage at the booting stage of japonica rice in cold region.【Method】The split plot design and the central combination test design were used to screen out the optimal application concentrations of CaCl2, glutamic acid(Glu), and γ-aminobutyric acid(GABA) through the membership function values of yield-related indicators under cold-water stress at booting stage.【Result】1) Under cold water stress at the booting stage, the optimal concentrations of exogenous CaCl2, Glu and GABA (CGG) were 1.87 mmol/L, 2.76 mg/L, 4.40 mmol/L, respectively and the average membership function value is the largest (0.932). 2) Compared with normal irrigation, the nitrogen photosynthetic efficiency of cold-region japonica rice decreased under cold stress at the booting stage. Compared with cold water stress at the booting stage, CGG application at the optimal concentration significantly increased the leaf area index (5.37%) and nitrogen photosynthetic efficiency (4.53%) of Dongnong 428 at the full heading stage; significantly increased leaf biomass per leaf area (2.56%) and leaf area index (3.39%) of Songjing 10 at the full heading stage. 3) Compared with cold water stress at the booting stage, the application of the optimal concentration of CGG increased the dry matter accumulation from heading stage to maturity in the cold-region japonica rice, but it was still lower than that of the normal irrigation, and there were differences between varieties and treatments. 4) Compared with cold water stress at booting stage, the yield and yield components of cold-region japonica rice increased with the application of the optimal concentration CGG, and the impact on cold-sensitive varieties was more significant. 5) Compared with the cold water stress at the booting stage, the average increase in temperature production efficiency of Dongnong 428 and Songjing 10 was 5.55% and 23.72% under the optimal concentrations of CGG, respectively. The contribution rate of exogenous CGG to the gap of temperature production efficiency is 8.99%.【Conclusion】The application of the optimal concentration of CGG under cold water stress at the booting stage promoted leaf growth of cold-region japonica rice at the full heading stage and increased the population nitrogen photosynthetic efficiency, which was beneficial to dry matter accumulation and yield formation after heading stage, thereby reducing the loss of cold-region japonica rice under cold water stress at booting stage and improving the temperature production efficiency of cold-region japonica rice.     

基于重组酶聚合酶扩增技术的玉米矮花叶病毒快速检测方法的建立

玉米Zea mays L.是重要的粮饲兼用作物。重要检疫性病毒玉米矮花叶病毒maize dwarf mosaic virus(MDMV)一直威胁玉米生产,为预防外来有害生物入侵,本研究以玉米可能携带的MDMV为目标,根据MDMV外壳蛋白(coat protein,CP)保守基因序列,基于重组酶聚合酶扩增技术(recombinase polymerase amplification,RPA),设计了3对RPA引物,筛选并优化了反应体系,建立了MDMV的快速检测方法,最终的RPA反应体系中引物终浓度为0.4 μmol/L,反应温度为42℃,反应时间为20 min。该方法可特异性检测MDMV,对MDMV CP阳性质粒样品的检测灵敏度可以达到105拷贝/μL(≈369 fg/μL),高于RT-PCR的检测灵敏度。该方法具有快速准确等优点,可用于玉米种子或植株中携带MDMV的检测。     

籼稻背景下抑制不同ALK等位基因表达对稻米品质的影响

【目的】稻米糊化温度是影响稻米品质的重要指标，该性状受主效基因ALK/SSII-3调控，ALK基因具有多个复等位基因，本研究旨在通过RNAi技术明确籼稻亚种中两个不同ALK等位基因的效应。【方法】以分别含有ALKc和ALKb等位基因的高糊化温度品种珍汕97B和低糊化温度品种龙特甫B为试验材料，使用RNAi技术构建ALK表达下调的转基因株系，通过对其稻米理化品质的测定来明确不同等位基因表达下调对稻米品质的影响。【结果】对不同转基因水稻目的基因的表达分析显示本研究中转基因株系的ALK基因受到了不同程度的干扰。重点分析了不同RNAi株系稻米的糊化温度，结果表明珍汕97B的RNAi转基因稻米的糊化温度极显著降低，而在低糊化温度品种龙特甫B背景中下调表达ALK基因后对糊化温度的影响较小；转基因株系与未转化亲本相比，米粉的起始糊化温度都显著降低，表现为提前糊化；在珍汕97B背景下干扰系的峰值温度与未转化亲本相比极显著降低，而在龙特甫背景下米粉的峰值温度与未转化对照相比显著降低。对不同转基因系的理化品质分析表明，ALK下调表达植株稻米的表观直链淀粉含量显著增加，下调表达ALK后会引起米粉峰值黏度和崩解值的改变。高糊化温度品种珍汕97B干扰系与未转化对照相比胶稠度呈现极显著性差异，而低糊化温度品种龙特甫干扰系的胶稠度与未转化对照相比没有差异。【结论】下调表达ALK等位基因对稻米理化品质产生显著影响，并且干扰不同等位基因的效应存在明显差异，即籼稻中的两个ALK等位基因的效应存在显著差异。