培养液NO3－/NH4+比对甜菜幼苗NO3－、NH4+吸收特性的影响

采用标准偏高糖型甜研7号与标准偏丰产型甜研8号2个二倍体纯系品种及水培方法，研究了子叶期（11 d）甜菜对NO₃^－和NH₄⁺ 的吸收特性以及不同NO₃^－/NH₄⁺ 比对苗期（31 d）甜菜吸收特性的影响。发现了子叶期甜研7号较甜研8号对NH₄^＋有较大的Vmax，有利于NH₄⁺的吸收。低NH₄⁺浓度比促进甜菜对NO₃^－的吸收。而且甜研7号受到的影响相对大于甜研8号。不同NO₃^－/NH₄⁺ 也影响甜菜对NH₄⁺ 的吸收速率，2品种所受的影响并不相同。2品种遗传特性不同导致了甜研7号对NH₄⁺ 的吸收较甜研8号敏感。高浓度NH₄⁺ 的存在促进了甜研7号与甜研8号对NH₄⁺ 的吸收。说明可以通过调节甜菜对NO₃^－与NH₄⁺ 的吸收与同化关系来调控甜菜的氮代谢，以提高甜菜的产量与质量。     

转根癌农杆菌介导的AtNHX1基因马铃薯的获得

构建重组表达质粒pBI12135-GZ+AtNHX1（带有CaMV 35S启动子），通过农杆菌将其携带的液泡膜Na⁺/H⁺逆向转运蛋白基因（AtNHX）转化马铃薯栽培品种“甘农薯2号”试管薯薄片和“克新2号”茎段。经根癌农杆菌侵染和共培养后，用50 mg L^-1卡那霉素 +300 mg L^-1头孢霉素筛选抗性芽，试管薯薄片获得30株抗性植株，抗性植株再生率为37%；茎段未获得抗性植株。抗性植株的总DNA用AtNHX1基因的特异性引物进行PCR检测，结果27株为阳性，占90%。Southern杂交结果证实，外源基因多以双拷贝整合到马铃薯的基因组中。Northern杂交结果表明，转基因植株可以进行AtNHX1基因mRNA的正常转录，但植株间存在着转录量的差异。该研究为耐盐马铃薯的培育奠定了良好的基础。     

紫花苜蓿Na+/H+ 逆向转运蛋白基因在拟南芥中表达提高转基因植株的耐盐性

以紫花苜蓿(Medicago sativa)为材料, 利用反转录PCR方法分离了NHX1全长cDNA(命名为MsNHX1)。Southern杂交结果表明, 在紫花苜蓿中存在一个小的液泡型Na⁺/H⁺逆向转运蛋白基因家族。序列分析表明, 该基因所编码的蛋白与拟南芥、水稻和棉花中液泡型Na⁺/H⁺逆向转运蛋白具有较高的同源性。在洋葱表皮细胞中瞬时表达MsNHX1-GFP融合基因的结果表明, MsNHX1定位在液泡膜上。Northern杂交发现该基因的表达受高浓度NaCl诱导。MsNHX1在盐敏感酵母突变体中表达可以提高转化子对NaCl的耐受性, 说明MsNHX1具有转运Na⁺的功能。在拟南芥中表达MsNHX1能显著提高植株耐受盐胁迫的能力; 而且在受到盐胁迫时, 转基因植株比野生型的渗透调节能力更强, 生物膜受破坏程度降低, 光合能力增强。以上研究结果表明MsNHX1是一个液泡膜Na⁺/H⁺逆向转运蛋白, 在植物耐受盐胁迫过程中起重要作用。     

转金属硫蛋白基因(MT1)烟草抗Cd2+胁迫的生理特性分析

将柽柳（Tamarix sp.）金属硫蛋白基因（MT₁，GenBank登录号：AB298390）插入植物表达载体pBI121，取代花椰菜病毒35S启动子下游的gus基因，利用农杆菌（Agrobactrium tumefaciens）介导法将MT1导入烟草（Nicotiana tobacum）基因组。对具有卡那霉素抗性，且经PCR-Southern检测和Northern杂交表现阳性的转基因株系进行抗Cd²⁺分析表明，金属硫蛋白基因的过量表达提高了转基因植株的抗Cd²⁺能力，在含200 µmol L^-1和400 µmol L^-1 Cd²⁺的MS培养基上，转基因植株的株高和鲜重均明显优于非转基因株系；在生理性状上表现为转基因植株MDA含量及POD活性明显低于非转基因株系，叶绿素含量和SOD活性比非转基因株系明显增加。     

辐射育种获得油菜（Brassica napus）高油酸材料

本研究用8~10万伦琴⁶⁰Coγ射线辐射双低油菜（B. napus）湘油15干种子，并对辐射后代进行高油酸连续选择，结果M₂、M₃、M₄油酸含量有不同程度提高，至M5油酸含量迅速提高，多数植株油酸含量在70%以上，最高油酸含量达93.5%。对高油酸突变体M604-855（种子油酸含量为91.     

利用回交导入系群体发掘水稻种质资源中的有利耐盐QTL

以中等感盐籼稻IR64与粳稻Tarom molaii培育的85个BC₂F₈回交导入系为材料，定位苗期在140 mmol L^-1 NaCl胁迫下影响叶片盐害级别、幼苗存活天数、地上部和根部的K⁺、Na⁺浓度等6个耐盐相关性状的QTL。幼苗存活天数与地上部Na⁺浓度呈极显著负相关，与地上部K⁺浓度呈显著正相关，与根部K⁺、Na⁺浓度无关，表明叶片盐害是由于地上部Na⁺积累过多造成的。根部K⁺浓度与Na⁺浓度高度正相关，但与地上部的K⁺、Na⁺浓度均无关，表明根对K⁺、Na⁺的离子吸收与向地上部运输存在不同的机制。检测到影响6个耐盐相关性状的23个QTL，包括影响叶片盐害级别的5个、幼苗存活天数的6个、地上部K⁺浓度的4个、地上部Na+浓度的4个、根部K+浓度的1个和根部Na+浓度的3个。影响地上部K⁺、Na⁺浓度与影响根部K⁺、Na⁺浓度的QTL分布在不同基因组区域，进一步表明根和茎对K⁺、Na⁺的吸收存在不同的遗传机制。通过比较图谱，发现影响耐盐相关性状的23个QTL中有12个（占52.2%）与以往不同群体中影响耐盐相关性状的QTL定位在同一或相邻的染色体区域。其中在第2染色体RM240~RM112区间检测到1个影响地上部所有4个耐盐相关性状的主效QTL，其增加耐盐性的有利基因来自供体Tarom molaii，适宜用作标记辅助选择耐盐性的遗传改良。对从种质资源中发掘“隐蔽”耐盐QTL进行了讨论。     

水稻(Oryza sativa)新型液泡Na+/H+逆向转运蛋白基因的特征分析和表达

以拟南芥AtNHX1 cDNA 片段作为探针，筛查水稻盐胁迫植株叶片cDNA 文库，获得与AtNHX1同源的水稻新型液泡Na⁺/H⁺逆向转运蛋白基因（OsANT1）。序列分析表明，OsANT1 全长cDNA为2 178 bp，包括一个长度为1 608 bp的完整开放阅读框，编码535个氨基酸残基。在DNA水平上，OsANT1基因含有15个外显子和14个内含子，长度为4 835 bp。OsANT1含有12个跨膜域，系统进化树分析结果表明，与来自拟南芥、水稻、小麦、玉米、大麦、马蔺和芦苇等的Na⁺/H⁺逆向转运蛋白高度同源。盐胁迫条件下，OsANT1的表达具有盐分诱导特征，且随着胁迫的增大而增加。表明该基因可能在水稻抵御盐分胁迫的过程中具有一定作用。     

十字花科黑腐病菌hrpW基因导入油菜之研究初报

为了研究十字花科植物黑腐病菌（Xanthomonas campestris pathovar campestris,简称Xcc）hrpW基因的功能及Xcc的致病机理,实验以甘蓝型油菜为试验材料,以5~6d无菌苗的带柄子叶为外植体,建立了高效稳定的带柄子叶再生体系,同时探索了影响根癌农杆菌介导转化甘蓝型油菜子叶柄的各种因素;并用农杆菌介导法将hrpW基因导入甘蓝型油菜中,经PCR检测法分析,证明hrpW基因已整合到油菜基因组中。hrpW基因编码HarpinXcc（十字花科植物黑腐病菌Harpin蛋白）,该转基因植株的获得,为进一步研究十字花科植物黑腐病菌的致病机理提供了材料,也为选育高效抗病油菜种质奠定了基础。     

水稻株高性状对大气CO2浓度升高的响应

以粳稻品种Asominori与籼稻品种IR24的杂交组合所衍生的染色体片段置换系（CSSLs）为材料，田间试验分别在FACE（CO₂浓度约570 µmol mol^-1）和对照（CO₂浓度约370 µmol mol^-1）下，对水稻株高性状的数量性状位点（QTL）进行了分析。结果表明，Asominori和IR24的株高、穗长、上位第一节间长和上位第二节间长在FACE和对照下的差异达显著水平；供试株系的4个株高性状对CO₂浓度升高都呈正负两种响应，其变化最大的株系为AI7和AI44（株高分别增加14.2 cm和降低4.54 cm），AI9和AI12（穗长分别增加3.56 cm和降低2.39 cm），AI39和AI27（上位第一节间长分别增加15.74 cm和降低1.49 cm），AI32和AI53（上位第二节间长分别增加8.09 cm和降低3.00 cm）；FACE和对照下分别检测出14和15个QTL，分布在除第2、7、9和第10号染色体外的各染色体上，其中5个（qPH6^-4、qPH8^-4、qPL8^-4、qPL12^-4和qLFN6^-4）在FACE和对照条件下同时检测到，分布在第6、8和第12染色体上，而其余的只在FACE或对照下检测到。这29个QTLs中，3个（qPH6^-4QE、qPH8^-4QE和qLSN5^-4QE）具显著的基因型与环境互作。在不同的CO₂环境下，测试性状发生不同程度的表型变异。结果推论，对CO₂浓度增加敏感的QTL位点，可能受到CO₂浓度增加的诱导，可见控制水稻株高性状的QTL与CO₂增加的环境发生了互作效应。     

高温胁迫对早稻根系质膜ATPase活性及NH4+吸收的影响

选用高温耐性品种农大228、082和高温敏感品种茉莉占、协青早B，比较研究了高温胁迫对根系质膜ATPase活性和NH₄⁺吸收速率的影响。结果表明，高温胁迫下，农大228和082根系质膜H⁺-ATPase和Ca²⁺-ATPase活性均比对照提高，而茉莉占和协青早B分别比对照大幅度下降，品种间差异显著(Duncan’s法检验)。根系质膜蛋白含量在高温胁迫后均表现下降。082的NH₄⁺吸收速率不受高温影响，热敏感型品种NH4+吸收速率受高温胁迫明显下降。     

含NPTⅡ标记基因的转基因抗虫棉室内快速鉴定方法

通过试验研究,提出了3种室内快速筛选和鉴定含NPTⅡ标记基因的转基因抗虫棉的方法.一是待检测棉子去种皮,在含卡那霉素培养基培养,根据幼苗子叶颜色和棉苗状态来辨别是否是转基因材料,卡那霉素最适浓度为0.75 g·L-1;二是去皮种子培养在无卡那霉素的萌苗培养基上,在棉苗子叶上涂抹卡那霉素溶液进行鉴定,最适浓度为4.0 g·L-1;三是在待测棉苗子叶上打孔、滴加一定浓度的卡那霉素,根据打孔并滴加卡那霉素部位的叶片颜色变化,鉴别并统计转基因植株,其最适浓度为2.0 g·L-1.3种方法均可在卡那霉素处理后4～7 d内快速鉴定出转基因植株.同时,对转基因阳性植株进行PCR鉴定,结果表明3种卡那霉素方法鉴定的准确率均在90%以上.     

转IPT基因油菜提高抗蚜虫能力

本研究利用农杆菌介导IPT(isopentenyl-transferases,异戊烯基转移酶)基因转化喀斯特特有甘蓝型油菜自交品种S65,在无激素的筛选培养基中获得再生植株14株,PCR检测12株外源基因整合入油菜基因组中.对其中5株转基因植株及5株野生型植株人工接种蚜虫试验,研究超量表达IPT基因油菜对蚜虫的抗性.结...     

不同激素浓度对橡胶草不同器官直接分化再生苗的影响研究

分别以橡胶草(Taraxacum kok-saghyz Rodin)叶片、叶柄、根段为外植体,以MS为基本培养基,比较不同激素浓度对橡胶草不同器官直接分化再生苗的影响。结果表明：橡胶草不同器官直接分化再生苗所需最佳激素浓度不同。(1)最适合诱导叶片和叶柄直接芽分化再生苗的激素浓度均为NAA 0.2 mg/L 6-BA 0.5 mg/L,其芽分化率和每外植体平均芽数分别为36.44%、38.83个和95.83%、10.56个;(2)最适合诱导根段直接芽分化再生苗的激素浓度为NAA 1.0 mg/L 6-BA 0.1 mg/L,其芽分化率和每外植体平均芽数分别为92.25%和21.49个。该研究为橡胶草高频再生体系的建立及进一步利用基因工程进行品种改良奠定了基础。     

Genetic analysis of new short petiole gene in soybean

The short petiole trait is valuable for the development of plant ideotype with high yield. Soybean breeding line, SS98206SP, showed extremely short petioles in greenhouse and field. In this study, the short petiole of two mutant lines, SS98206SP and D76-1609, were investigated to determine the genetic segregations. These two mutants were crossed with each other and with two normal petiole genotypes. Genetic analysis indicated that the short petioles in D76-1609 and SS98206SP were controlled by a single recessive gene, respectively. The short petiole gene in SS98206SP was non-allelic with lps, conferring short petiole in D76-1609. Two recessive genes showed complementary relationship having short petioles with recessive homozygote (LPS1-lps?lps?, lps1lps1LPS?-, lps1lps1lps?lps?). Our data indicated that the short petioles in SS98206SP were controlled by a single recessive gene designated as lps3.     

Outcrossing frequencies from multiple high erucic acid oilseed rape fields to a central receptor field

Genetically modified oilseed rape is currently grown on about 23 % of the global oilseed rape acreage. In order to separate transgenic and non-transgenic oilseed rape production and to ensure co-existence of different agricultural cultivation schemes, as is specified by the European legislation, confinement measures have to be defined. Pollen-mediated gene flow is the most important means by which transgenes are dispersed between fields. In contrast to the majority of the previous investigations the objective of this study was to assess the extent of gene flow in the case of multiple pollen-donor fields. A high erucic acid rape genotype was used as biochemical marker for the quantification of outcrossing into a low erucic acid oilseed rape variety. Outcrossing data were obtained from two experimental locations. As expected, multiple pollen sources in a fragmented landscape can result in high gene transfer frequencies, thus requiring larger isolation distances than a field design with a single pollen donor source. The results of the study are transferable to homozygous transgenic oilseed rape varieties.     

基于生命周期法的中国2004—2015年油菜生产氮足迹分析

The loss of reactive nitrogen in agriculture has increasingly become serious, systematic analysis of reactive nitrogen emissions from agricultural production is conducive to promoting the green and healthy development of agriculture in China. Based on the statistical data of crop yield and farmland investment in the oilseed rape production of China, the spatiotemporal dynamic change of nitrogen footprint (NF) and its composition of oilseed rape during 2004-2015 in China was estimated using the theory of NF and life cycle assessment method in the agricultural sector. The NF of oilseed rape was 7572.0 g N-eq ha ^-1 during 2004-2015, and decreasing year by year, with an average annual decline of 1.0%. The main components of the NF were from fertilizer application (20%) and NH₃ emission (54%) in the oilseed rape production. Obvious differences also were observed among mainly agricultural provinces in China, that is, the higher nitrogen footprint per area (NF_a) and nitrogen footprint per yield (NF_y) in Inner Mongolia, Shanghai, and Jiangsu region. The NF_a and NF_y in high yield region were significantly higher than those in low yield region. Among them, the NF of nitrogenous fertilizer, phosphate fertilizer and compound fertilizer increased by 80.6%, 76.9%, and 57.8%, respectively (P < 0.05). The NF_a of rapeseed in China showed a trend of increase with the increase of rapeseed yield, but there was no significant correlation. The results suggest that improving crop management practices that limit fertilizer consumption is an important measure for the green and healthy development of rapeseed in China.     

芽孢杆菌Bam22对油菜根肿病的防治效果研究

由芸薹根肿菌(Plasmodiophora brassicae)侵染引起的根肿病是限制油菜生产的主要病害之一,苗期即可发病,目前该病害的主要防治措施有选用抗病品种和使用化学药剂,但农药残留问题日益突出,故提出采用生物防治的策略防控油菜根肿病。笔者采用拌种与苗期灌根的方式研究了芽孢杆菌(Bacillus Bam22)对油菜根肿病的田间防治效果,苗期调查结果表明芽孢杆菌Bam22拌种处理(T1)和芽孢杆菌Bam22+高岭土拌种处理(T2)较清水对照(CK)的发病率降低了3.4%~20%,防效达44%~52%;T1、T2与药剂对照氰霜唑拌种处理(T3)的株高和发病率无明显差异,但防效提高20%~40%。花期调查结果表明T1和T2处理较清水对照(CK)的发病率降低了10%~13%,防效介于28%~51%;T1、T2处理较药剂对照(T3)的发病率降低了6.67%~10%,防治效果提高了5%~22%,单株产量增加了23.97%~34.29%。以上研究结果表明,采用芽孢杆菌Bam22拌种结合苗期灌根,能够有效降低油菜苗期和花期根肿病的发病率,显著降低油菜的产量损失率。芽孢杆菌Bam22对油菜根肿病有较好的控制效果,具有开发成为生物农药的潜力。     

氮肥和隔根对胡麻/大豆间作体系种间关系及间作优势的调控效应

为合理利用氮肥和隔根模式技术优化间作作物种间关系,进一步发挥种间优势提供理论依据,以胡麻间作大豆为研究对象,采用盆栽试验,研究不施氮(N0)和施¹⁵N 80 mg/kg(N80),根系完全分离(T1)、根系不分离(T2)、根系尼龙网分离(T3)3种隔根模式对胡麻/大豆间作优劣势和种间竞争力的影响。结果表明,间作胡麻干物质积累量在各处理间无显著差异,间作大豆全生育期干物质积累量表现为N80显著高于N0处理。间作胡麻单株有效蒴果数、每果粒数、千粒重和籽粒产量均在N80T2处理下最大,其中,籽粒产量较其他处理显著高出41.94%~137.84%。间作大豆根瘤数为N0>N80,单株结荚数、每荚粒数和百粒重均为N80T2处理最大,籽粒产量则为N80T3处理最大,但与N80T2处理差异不显著,较其他处理显著高出4.33%~13.99%。所有处理的土地当量比(LER)均大于2,N80T2时最大。不同施氮模式下,T2处理的种间竞争力表现为胡麻强于大豆(A_F>0,CR_F>1),T3处理则恰好相反(A_F<0,CR_F<1)。说明胡麻/大豆间作体系中施¹⁵N 80 mg/kg、根系不分离有利于协调胡麻和大豆种间关系,促进作物增产。     

Characterization of Brassica nigra chromosomes and of blackleg resistance in B. napus–B. nigra addition lines

Brassica napus-B. nigra addition lines were previously created using the variety ‘Darmor’ as the oilseed rape genetic background. Two isozyme loci and 46 RAPD markers were added on five different B. nigra chromosomes. The oilseed rape variety used was highly susceptible to blackleg at the cotyledon stage and only the addition of chromosome 4 gave the same level of blackleg resistance as B. nigra. This resistance was efficient whatever the isolates used. A significant effect on the development of stem canker under field conditions was observed only for the line carrying chromosome 4 which was more resistant than the susceptible control. The potential effects of two other chromosomes have to be confirmed. F1 hybrids obtained by crosses between two highly susceptible lines and the monosomic addition line carrying chromosome 4 were examined under field conditions. No effect of the oilseed rape genetic background on the expression of resistance was detected. The introduction of this resistance and mapping of the gene(s) into oilseed rape varieties are discussed.     

Genetic diversity in mustard (Brassica juncea L.) germplasm from Pakistan as determined by RAPDs

The genetic diversity and the relationships among a collection of mustard (B. juncea) germplasm, including 41 accessions collected from Pakistan, 6 oilseed cultivars/ lines and 5 Japanese vegetable cultivars, were evaluated using random amplified polymorphic DNA (RAPD) markers. A total of 198 polymorphic amplified products were obtained from 30 decamer primers. Of these, 14 were unique to the accession PAK-85835 and 37 were specific to PAK-85839. Based on pair-wise comparisons of RAPD amplification products, genetic similarity was estimated using similarity coefficients of Nei & Li (1979) and a dendrogram was constructed using the unweighted pair-group method with arithmetic averages (UPGMA). Cluster analysis based on these genetic similarities placed most of the collected germplasm and oilseed cultivars/lines close to each other, showing a low level of polymorphism between the oilseed accessions collected in Pakistan. However, the clusters formed by the oilseed collections and cultivars were distinct from those formed by the vegetable cultivars. A low level of genetic variability of oilseed mustard in Pakistan was attributed to the selection for similar traits and horticultural uses. The farmers' preference for more remunerative crops and perhaps the close parentage of these accessions further contributed towards their little diversity. The study demonstrated that the RAPD is a simple and fast technique to compare the genetic relationships and the patterns of variation among accessions of this crop. This revised version was published online in August 2006 with corrections to the Cover Date.