玉米抗南方型锈病基因共分离分子标记的研究

本文以玉米(Zea mays L.)抗南方型锈病自交系P₂₅和感病自交系F₃₄₉的F₁与F₃₄₉回交,并连续回交所得BC₅代的F₃₄₉抗病近等基因系（nearly isogenic lines,NILs）为材料,以RGA（resistance gene analogs）的方法,在抗病自交系中克隆出一条321 bp的特异性DNA条带。根据序列比对的结果,设计出新的引物作为分子标记,在96株BC₇、BC₄F₅NILs及其亲本和杂种F₁群体上进行鉴定,其中94株的田间抗感结果与分子检测结果一致,选择的有效率达97.9%。此标记扩增效果清晰,可重复性强,在抗玉米南方型锈病的分子标记辅助育种研究中有应用价值。     

The Relationship Between Developmental Accumulation of Leaf Soluble Proteins and Vernalization Response of Wheat （Triticum aestivum L.em. Thell）

The relationship between vernalization requirement and quantitative and qualitative changes in total leaf soluble proteins were determined in one spring （cv. Kohdasht） and two winter （cvs. Sardari and Norstar） cultivars of wheat （Triticum aestivum L.） exposed to 4℃. Plants were sampled on days 2, 14, 21 and 35 of exposure to 4℃. The final leaf number （FLN） was determined throughout the vernalization periods （0, 7, 14, 24, and 35 d） at 4℃. The final leaf number decreased until days 24 and 35 in Sardari and Norstar eultivars, respectively, indicating the vernalization saturation at these times. No clear changes were detected in the final leaf number of Kohdash cultivar, verifying no vernalization requirement for this spring wheat cultivar. Comparing with control, clear cold-induced 2-fold increases in proteins quantity occurred after 48 h following the 4℃-treatment in the leaves of the both winter wheat cultivars but, such response was not detected in the spring cultivar. However, the electrophoretic protein patterns showed between-cultivar and between-temperature treatment differences. With increasing exposure time to 4℃, the winter cultivars tended to produce more HMW polypeptides than the spring cultivar. Similar proteins were induced in both Sardari and Norstar winter wheat cultivars, however, the long vernalization requirement in Norstar resulted in high level and longer duration of expression of cold-induced proteins compared to Sardari with a short vernalization requirement. These observations indicate that vernalization response regulates the expression of low temperature （LT） tolerance proteins and determines the duration of expression of LT- induced proteins.     

Precision phenotyping of imidazolinone-induced chlorosis in sunflower

Chlorosis level is a useful parameter to assess imidazolinone resistance in sunflower (Helianthus annuus L.). The aim of this study was to quantify chlorosis through two different methods in sunflower plantlets treated with imazapyr. The genotypes used in this study were two inbred lines reported to be different in their resistance to imidazolinones. Chlorosis was evaluated by spectrophotometrical quantification of photosynthetic leaf pigments and by a bioinformatics-based color analysis. A protocol for pigment extraction was presented which improved pigment stability. Chlorophyll amount decreased significantly when both genotypes were treated with 10 μM of imazapyr. Leaf color was characterized using Tomato Analyzer^® color test software. A significant positive correlation between color reduction and chlorophyll concentration was found. It suggests that leaf color measurement could be an accurate method to estimate chlorosis and infer chlorophyll levels in sunflower plants. These results highlight a strong relationship between imidazolinone-induced chlorosis and variations in leaf color and in chlorophyll concentration. Both methods are quantitative, rapid, simple, and reproducible. Thus, they could be useful tools for phenotyping and screening large number of plants when breeding for imidazolinone resistance in this species.     

玉米pTAC2影响苗期叶片叶绿素合成的转录组分析

【目的】叶绿素是参与光合途径最为重要的光合色素。叶绿体的发育及叶绿素的合成在很大程度上依赖于质体基因组与核基因组之间的双向信号传导来精确协调基因表达。通过对白化表型的CRISPR/Cas9-ZmpTAC2转基因阳性纯合突变材料进行RNA-seq研究,筛选和鉴定参与叶绿素合成的相关基因,为明确叶绿素的合成途径奠定基础。【方法】以CRISPR/Cas9-ZmpTAC2玉米转基因编辑纯合突变株系为研究材料,使用透射电镜观察叶绿体超微结构和分光光度法测定叶片叶绿素含量,确定叶绿体发育状态及叶绿素合成情况。对转基因阴性材料(CK)和CRISPR/Cas9-ZmpTAC2转基因纯合编辑材料(zmptac2)苗期叶片取样进行转录组测序。通过生物信息学分析,寻找CK与zmptac2间差异表达的基因;qRT-PCR对差异表达基因进行验证。通过酵母双杂交筛选与玉米pTAC2互作的蛋白质。【结果】共获得15株T0转基因植株,包括绿色植株(7株)和白色植株(8株)。绿色幼苗中3株为转基因阴性材料,4株为转基因阳性(2株为未编辑,2株为杂合编辑突变),白色植株(8株)均为转基因阳性纯合编辑。与CK相比,突变体(...     

鹰嘴豆LHY基因cDNA克隆及生物信息学分析

从鹰嘴豆中克隆生物节律钟LHY基因的cDNA全长序列,进行序列信息学分析。通过同源克隆策略,利用RT-PCR技术获得核心片段,结合5＇-RACE和3＇-RACE技术,克隆得到鹰嘴豆生物节律钟基因LHY的cDNA全长序列,其核苷酸序列长度为3 061 bp,包括2 220 bp的完整开放阅读框（ORF）,编码739个氨基酸。验证后命名为CarLHY基因,获得基因登录号为KJ558378。生物信息学研究表明CarLHY基因cDNA序列与其他植物LHY基因具有较高的相似性;预测CarLHY蛋白不具有跨膜结构;为转录因子,定位于细胞核中;不具备信号肽。对CarLHY蛋白功能结构域预测表明,蛋白质核心结构存在符合转录因子与DNA结合的常见功能域HTH。蛋白系统进化树显示,与大豆分子进化距离最近,其次是黑杨、拟南芥。     

苦楝果实提取物对几种芽孢杆菌的抑菌活性

将采摘的苦楝果实烘干、粉碎后,以甲醇为溶剂,用超声波法提取后,配成不同浓度的苦楝果实提取液,测定其对巨大芽孢杆菌、短芽孢杆菌、地衣芽孢杆菌和枯草芽孢杆菌4种芽孢杆菌的抑菌圈直径和抑菌百分率。结果表明:苦楝果实提取液浓度越高,其抑菌作用越强。苦楝果实提取液对枯草芽孢杆菌的抑菌活性最大,对短芽孢杆菌的次之,对巨大芽孢杆菌和地衣芽孢杆菌的抑菌活性最小。     

中国海区不同养殖品系海带与长海带之间的遗传关系

对中国海区海带(Laminaria japonica)不同栽培品系(种)及长海带(L.longissirna)的配子体无性繁殖系进行12个酶系统的同工酶检测,获得了31个基因位点,酶谱分析结果说明,来自同一株海带的雌、雄配子体无性繁殖系具有较高的遗传相似性,但“860”品系可能因杂合度大,导致该品系雌、雄配子体无性繁殖之间的遗传距离较远。对它们相应的亲本孢子体进行RAPD分析,30个随机引物共得到185个扩增位点,其中多态性位点76个,占41％。利用UPGMA对同工酶及RAPD所得到的遗传距离进行聚类分析,表明中国海区养殖海带的不同品系或品种之间的遗传相似性较大,而“烟杂1号”与长海带之间具有较近的亲缘关系。基于同工酶和RAPD图谱的结果推测,“烟杂1号”是一个新的杂交种,其母本可能是“荣杂1号”,而父本可能是长海带。     

AMF对紫花苜蓿抗盐生理特性的影响

试验选用3种丛枝菌根真菌(AMF)接种苜蓿品种金皇后,研究了5个梯度NaCl(0、50、100、150和200 mmol/L)对金皇后苜蓿抗盐生理特性的影响。结果表明:金皇后苜蓿的脯氨酸含量和可溶性糖含量随着NaCl浓度的增加总体呈先升高后降低的趋势,在100 mmol/L NaCl处理下达到最高,接种Gm真菌的二者含量分别为6.37和1.91μg/g;金皇后丙二醛(MDA)含量随着NaCl浓度的增加呈升高趋势,且在4个NaCl浓度之间差异显著;超氧化物歧化酶(SOD)和过氧化物酶(POD)活性随着NaCl浓度的增加呈先升高后降低的趋势,在150 mmol/L NaCl处理下二者活性下降,接种Gm真菌的2种抗氧化酶活性分别比处理组下降了48.48%和43.71%;同时金皇后在盐环境下光合指标的变化幅度较大,其中Ci随着NaCl浓度的增加而增加,P_n,G_s和T_r随着NaCl浓度的增加而下降;金皇后最大耐盐程度为150 mmol/L NaCl,且接种摩西球囊霉(Gm)效果优于幼套球囊酶(Ge)和根内球囊霉(Gi)。     

Improvement in the Adaptation of Lygeum Spartum L. to Salinity In the Presence of Calcium

Lygeum spartum L. has been recently introduced in areas where salinity is high in soils. However, there are no studies about the physiological response of these plants to salt excess. The effect of sodium chloride (NaCl) on plant growth and water status was studied. Also, the effect of calcium (Ca) addition to salinity conditions was analyzed because of the coexistence of salinity and calcareous soils. Dry weight (DW), transpiration, and osmotic potential (Ψπ) decreased with elevated NaCl and were restored with Ca²⁺, whereas moderate salinity had no effect. Fresh weight (FW), water potential (Ψω), and root hydraulic conductance (L ₀) decreased with salinity; Ca²⁺ supply had an ameliorative effect at moderate salinity. Sodium (Na⁺) increased in leaf sap at high levels of NaCl and was decreased by Ca²⁺. Lygeum spartum showed a resistance to moderate salinity, but the effect of Ca²⁺ depends on salinity intensity. Thus, the role of Ca²⁺ in the tolerance to salinity was emphasized.     

扁蓄提取物对小菜蛾生物活性的影响

研究扁蓄(Polygonum aviculare L.)乙醇提取物对小菜蛾(Plutella xylostella L.)触杀、胃毒、拒食及抑制生长的影响,比较扁蓄不同部位的活性,并对提取溶剂进行选择。结果表明,扁蓄乙醇提取物对小菜蛾的主要作用方式为触杀。