Morphological and chemical characteristics of doubled haploids of flue-cured tobacco combining resistance to Thielaviopsis basicola and TSWV

Thielaviopsis basicola and Tomato spotted wilt virus (TSWV) are the most important problems in a moderate climate zone. Previously obtained doubled haploids (DH) of F₁ hybrids of the flue-cured line WGL3 resistant to Th. basicola and the dark-cured line PW-834 carrying RTSW-al gene provided the research material. Biological tests and SCAR markers linked with TSWV were applied to confirm resistance of DH. Lines combining resistance to TSWV and Th. basicola were evaluated for morphological and chemical characteristics. Most of DH were significantly shorter than parents but two lines, 31/A/2 and 31/B/3, were close to the flue-cured WGL3. Usually DH possessed fewer leaves while one of them 31/B/3, exceeded parental forms. The doubled haploids flowered later than their parents. The most negative effect was reduced area of mid-position leaves of DH. It might be explained by a recombination during microsporogenesis in F₁, however the influence of ‘Polalta’-derived RTSW-al gene cannot be excluded. Extensive line to line variation for nicotine and sugars content was not associated with the genes for TSWV and Th. basicola resistance. Biological tests and field performance of DH revealed potential to overcome the negative effect of coupling between the RSTV-al gene and genes responsible for the morphological deformations.     

Construction of Recombinant Expression Plasmid pYELmleA of mleA Gene and Expression in Saccharomyces cerevisiae

苹果酸－乳酸酶是进行MLF的功能酶。笔者进行酒酒球菌SD-2a的苹果酸－乳酸酶基因重组表达质粒的构建，利用来自重组质粒pLmleA的mleA基因，以PGK1强启动子和ADH1终止子为调控元件，以大肠杆菌－酵母菌穿梭质粒YEp352为载体，构建了重组表达质粒pYELmleA，并转化酿酒酵母（Saccharomyces cerevisiae）YS58。酵母转化子用含有亮氨酸、组氨酸和色氨酸的YNB平板筛选鉴定。SDS-PAGE检测表明获得的转化子表达了约60KD的目标蛋白，斑点杂交检测表明目的基因mleA转化到     

冬小麦–夏玉米周年生产条件下夏玉米的适宜熟期与积温需求特性

本研究旨在探讨冬小麦–夏玉米周年生产条件下黄淮海区夏玉米的适宜熟期与积温需求特性。选用郑单958(ZD958)、先玉335(XY335)、登海605(DH605)、登海618(DH618)和登海661(DH661),设置5月21日、5月31日、6月10日和6月20日4个播期,研究表明,播期对夏玉米生理成熟所需积温无显著影响,各品种生理成熟所需要的积温主要取决于品种自身的特性,DH618、XY335、ZD958、DH605、DH661的生育期和生理成熟所需要积温分别为110、112、116、116、121 d和2800、2880、2945、2950、3025°C d。冬小麦-夏玉米周年生产条件下,夏玉米最大可能的生长期约107~112 d(自6月15日至10月1~5日),积温约2800°C d,难以满足现有品种的生产需要。夏玉米直播晚收、冬小麦适期晚播有利于周年产量提高,但目前广泛推广的夏玉米品种生育期过长(约120 d),适时晚收仍难以完全生理成熟,机收籽粒损伤严重。可见,冬小麦–夏玉米周年生产条件下夏玉米最大可能的生长期和有效积温不能满足目前广泛推广的夏玉米品种所需生育持续期和积温,且适时晚收仍难以完全生理成熟,黄淮海区亟需生育期适宜(生育期≤107 d)的高产夏玉米新品种。     

Simulations and practical problems of applying multiple marker assisted selection and doubled haploids to wheat breeding programs

Marker assisted selection (MAS) and wheat doubled haploids (DH) are relatively new technologies, recently applied to wheat breeding programs. Simulations demonstrate that DHs increase the efficiency of MAS, and offer faster strategies for combining large numbers of genes with a minimum number of marker tests. When small numbers of marked loci (1-3) are selected simultaneously, selection of DH progeny is 5-6 times more efficient than selecting F₄ derived families. Combining 4-8 marked loci, screening of F₂ plants and using only those plants homozygous or segregating for all of the marked loci as parents for DH production (10-31% of F₂ plants) is 3-10 times as efficient as using F₁ plants. A number of protocols have been proposed involving sib-matings and selection to fix some genes, with further selection in the second generation to improve the proportion of useful DH lines. In one scheme (recombinant F₂ selection) all F₂ plants, either homozygous or heterozygous for the marked alleles, are intercrossed at random and the recurrent F₁ plants still having these alleles are used for DH production. An alternative strategy (recurrent DH selection) is to select from an initial DH population and intercross those lines having most favourable marked loci with a second cycle of DHs to fix all favourable marked loci. Combining more than 12 marked gene loci does not seem feasible, due to the very large numbers of F₂s (>2000) required. This has implications when using MAS for quantitative trait loci, where many minor gene loci would have to be combined. Direct selection for some multi-genic quantitative traits amongst the DH lines may be more efficient than using MAS where recurrent selection is used. At the Cereal Research Centre, the practical problems of using these protocols as part of the spring wheat breeding program are being evaluated. This revised version was published online in August 2006 with corrections to the Cover Date.     

玉米Reid种质DH系遗传分析

为探讨玉米Reid种质遗传改良效果,本研究以Reid类群重要种质J1491、J1492、J1493、J1495、J1498与PH6WC构建基础群体,通过DH技术育成DH系,经鉴定从中选取5个代表DH系6DH6、6DH7、6DH9、6DH10、6DH12作母本,以Non-Reid群5个优良自交系作父本,按NCII设计组配25个杂交组合,对DH系单株产量及其构成因素的杂种优势、配合力和相关方面进行了研究。结果表明,单株产量杂种优势以组合6DH7×J1628最高为180.22%,组合6DH12×J1673最低为54.94%;单株产量一般配合力效应值最高的有6DH7和6DH10,穗长、穗粗亦为正效应,因此认为6DH7和6DH10更具应用潜力;特殊配合力效应值最高的组合是6DH7×J1673。各产量性状中穗长、穗粗和秃尖长,狭义遗传率高,主要以基因加性效应为主。本研究为鉴定有潜在价值的DH系及杂交种提供选择,为玉米遗传改良提供参考。     

Mapping of QTL associated with waterlogging tolerance and drought resistance during the seedling stage in oilseed rape (Brassica napus)

Soil waterlogging and drought are major environmental stresses that suppress rapeseed (Brassica napus) growth and yield. To identify quantitative trait loci (QTL) associated with waterlogging tolerance and drought resistance at the rapeseed seedling stage, we generated a doubled haploid (DH) population consisting of 150 DH lines from a cross between two B. napus lines, namely, line No2127-17 × 275B F₄ (waterlogging-tolerant and drought-resistant) and line Huyou15 × 5900 F₄ (waterlogging-sensitive and drought-sensitive). A genetic linkage map was constructed using 183 simple sequence repeat and 157 amplified fragment length polymorphism markers for the DH population. Phenotypic data were collected under waterlogging, drought and control conditions, respectively, in two experiments. Five traits (plant height, root length, shoot dry weight, root dry weight and total dry weight) were investigated. QTL associated with the five traits, waterlogging tolerance coefficient (WTC) and drought resistance coefficient (DRC) of all the traits were identified via composite interval mapping, respectively. A total of 28 QTL were resolved for the five traits under control conditions, 26 QTL for the traits under waterlogging stresses and 31 QTL for the traits under drought conditions. Eleven QTL were detected by the WTC, and 19 QTL related to DRC were identified. The results suggest that the genetic bases of both waterlogging tolerance and drought resistance are complex. Some of the QTL for waterlogging tolerance-related traits overlapped with QTL for drought resistance-related traits, indicating that the genetic bases of waterlogging tolerance and drought resistance in the DH population were related in some degree.     

Variation in Doubled Haploid Plants of Wheat Obtained through Wheat (Triticum aestivum) × Maize (Zea mays) Crosses

Variation was investigated in 110 doubled haploid (DH) lines of wheat derived from wheat × maize crosses. Field observation revealed visible variations in 15 DH2 lines such as extreme dwarfism, low seed fertility, alteration of spike type and stripes. Six agronomic traits, i.e., heading date, spike number/ plant, culm length, spike length, seed fertility and grain weight were statistically analyzed in the DH2 and DH3 generations. Out of the 88 DH2 lines/DH3 groups, 26 %/64 % showed significant differences from the parental variety in the means of one or more traits. Ranges of the DH3 lines were larger than those of the DH2 lines, except for spike number/plant. Furthermore, analyses of variance within and between DH lines showed the presence of heterogeneity/heterozygosity in the DH2 lines/plants. These results indicated the occurrence of gametoclonal variation in the DH lines. It is considered that most of the variations detected were due to the colchicine treatment rather than to the 2,4-D treatment or in vitro culture.     

Early diagnosis of ploidy status in doubled haploid production of maize by stomata length and flow cytometry measurements

Reliable discrimination of haploid (H), doubled haploid (DH) and crossing (C) plants in early growth stages could streamline DH production in maize. By detecting in early growth stages undesirable sterile H plants and undesirable heterozygous C plants, a large proportion of resources required for DH production could be saved. The goal of this study was to evaluate the effectiveness of early classification of plants in growth stages V3‐V4 as H, DH or C in the context of DH production using flow cytometry and stomata length. As the reference classification, we used a field score based on plant phenotype commonly applied in DH production and research. Our results show that identification of misclassified C seeds is possible because the overlap in distributions of stomata length between H&DH and C plants is small and the association between flow cytometry and the reference field score is high. In contrast, overlap between H and DH distributions is substantial. Consequently, the main application we see for these classification methods in early growth stages is the identification of C seedlings.     

不同马铃薯品种的氮利用效率及其分类研究

以7个马铃薯品种为供试材料,设置大田条件下施氮和不施氮2种处理,在对块茎产量和植株氮素吸收、利用评价的基础上,将不同氮效率品种马铃薯分类并解析了其差异机制。基于2016年施氮和不施氮条件下各品种马铃薯的平均产量,把不同氮效率品种马铃薯分为双高效型、低氮高效型、高氮高效型和双低效型。2017年选择双高效型、低氮高效型、双低效型的代表性品种,对各类型氮效率差异进一步解析表明,双高效型氮素利用效率显著高于另两个类型,氮素吸收效率则是双高效型、低氮高效型显著高于双低效型。不施氮条件下,双高效型马铃薯的干物质累积量在整个生育时期均显著高于另2个品种;双高效型、低氮高效型氮素累积速率在出苗后0～50d显著高于双低效型马铃薯。施氮条件下,双高效型马铃薯的干物质累积量显著高于另2个品种,与双低效型马铃薯相比,双高效型和低氮高效型产量的提高主要归因于它们前期较高的干物质累积;双高效型氮素累积速率显著高于双低效型、低氮高效型。双高效型马铃薯在各生育期的物质生产和氮素吸收能力强,从而有利于氮效率提升和产量的形成。该研究结果可为马铃薯氮高效品种筛选和利用提供理论支撑。     

同一基础材料的玉米双单倍体(DH)系配合力的分析

利用高频玉米单倍体诱导系,对选系材料M35/F35的F1代进行诱导获得大量单倍体籽粒,经自然加倍获得一批纯合双单倍体(DH)系,用测验系京24组配杂交组合分析DH系的配合力表现。结果表明,来源于同一基础材料的不同DH系之间,配合力差异较大,部分DH系的配合力比亲本增加,说明采用单倍体育种可以选育到高配合力的优良玉米自交系,组配出优良品种,从而加快育种进程,应用前景广阔。     

Development and characterization of an eggplant (<Emphasis Type="Italic">Solanum melongena</Emphasis>) doubled haploid population and a doubled haploid line with high androgenic response

We developed an eggplant doubled haploid (DH) population from a commercial hybrid through androgenesis in microspore culture. Morphological variation, reproductive ability and androgenic responsiveness were evaluated. The DH population showed segregation in vegetative traits related to leaf, flower and fruit, and in reproductive traits such as fruit and seed setting or germination rate. The DH population and subsequent generations also presented variation in the androgenic response, with null, low and high response lines. From this population, we were able to identify the first eggplant highly androgenic DH line (DH36), remarkably similar to the donor hybrid in terms of morphology and reproductive ability, but stably producing four times more calli than the hybrid. The segregating DH population is potentially useful for genetic studies and mapping of several traits, whereas the highly androgenic line DH36 may be used as a model line to facilitate the study of eggplant androgenesis and embryogenesis for both basic and applied research.     

辣椒DH群体果实性状的分离及与F2群体的比较

为分析DH群体的遗传稳定性及其DH群体内性状分离与同来源的F2群体之间异同，本研究利用辣椒花药培养技术，构建一个由“羊角椒(97403)×方灯笼甜椒(97410)”的杂交而成的牛角椒组合作为供体的且由103个DH系组成的DH群体，对该DH群体进行单果重等5个主要果实性状的遗传表现进行分析，并与同来源的F2群体进行比较。结果表明：原供体及其双亲的5个果实性状的变异系数均较小，三者之间的各性状均存在显著差异，说明原供体及其双亲的整齐一致性；对DH群体和F2群体各性状的平均值、变异系数及其性状分离区间的分析和对比结果表明，5个果实性状均是受多基因控制的数量性状，且由于基因重组无论是DH群体还是F2群体均能产生正向和负向两个方向的超亲基因型。但DH群体与F2群体比较各性状的分离区间明显增大，超亲分离类型明显增加，且获得的各DH系均为稳定遗传的自交系，提高了花培育种的选择效率。     

DH群体内株系对籼、粳亚种的亲和性

以窄叶青8号（ZYQ8）/京系17(JX17)的13个花培DH株系和培矮64／02428的11个花培DH株系为材料,分析了它们的籼粳分化及其对籼、粳亚种杂交的亲和性。结果表明,无论亲本是否具有广亲和性亲缘,两个DH群体均出现广亲和性良好的株系,且多数株系属籼粳中间类型。这说明虽然广亲和性是遗传的,但是籼粳基因的重组确实可以产生广     

Doubled-haploid versus single-seed descent and S1-family variation for testcross performance in a maize population

Progress made in the in situ gynogenesis technique since 1990 now allows production of a high number of maize (Zea mays L.) doubled-haploid (DH) lines. The aim of the study was to compare DH lines versus selfing lines for testcross performance. DH and single-seed descent (SSD) lines were produced from random S₁ progenies of a broad-base population. For grain yield, kernel moisture, plant height, ear height and leaf length, the three population means were similar. Except for kernel moisture, the genetic variance of DH lines was nearly twice as high as the genetic variance of S₁ families, as expected. On the other hand, genetic variance among SSD lines was only 1.5 times higher than the genetic variance of S₁ families. This lower variance could be due to a selection bias in the method of production of SSD lines. However, for all traits, heritability of SSD or DH lines was higher than heritability of S₁ families. Epistasis effects in DH progenies were not significant. The consequence was a high correlation between S₁ testcross progenies and DH or SSD testcross progenies, meaning that the S₁ testcross value can be used to select the best families from which DH lines will be extracted. As a whole, the observed variation in DH lines appeared to be more in accordance with the observed variation among S₁ families than with the observed variation among SSD lines.     

小麦–华山新麦草异代换系DH2322的分子细胞遗传学鉴定

利用分子标记、细胞学、基因组原位杂交(GISH)等技术,结合田间农艺性状调查,对小麦–华山新麦草七倍体材料H8911与硬粒小麦D4286杂交F4代分离群体中株系DH2322进行了综合鉴定。华山新麦草基因组特异SCAR标记鉴定表明,DH2322含有华山新麦草遗传物质;细胞遗传学观察显示,DH2322染色体构型为2n=42=21 II;有丝分裂和花粉母细胞减数分裂中期I基因组原位杂交(GISH)鉴定表明,DH2322的染色体由40条小麦染色体和2条华山新麦草Ns染色体构成,且2条Ns染色体能完全配对为一个二价体;SSR和STS分子标记分析表明,DH2322缺失小麦D基因组的2D染色体,而含有华山新麦草的2Ns染色体;农艺性状分析结果表明,DH2322具有双亲的形态学特征,结实性好,穗长和穗粒数显著大于亲本。     

龙眼采后腐烂相关细菌与真菌的分离与鉴定

采用稀释涂布平板法,从龙眼腐烂果实中分离细菌和真菌,并结合菌落形态观察以及16S rDNA和ITS序列分析对所分离的细菌和真菌进行鉴定。结果表明,采用稀释涂布平板法,从龙眼腐烂果实中分离得到11株细菌和7株真菌。菌落形态观察以及16S rDNA和ITS序列分析结果表明,所分离的细菌主要鉴定为奥斯陆莫拉菌、短小芽胞杆菌、柠檬明串珠菌、肠膜状明串珠菌和东方醋酸菌。所分离的真菌主要鉴定为淡色生赤壳菌、葡萄座腔菌科真菌、间座壳菌、疖葡萄座腔菌和木贼镰孢菌。从龙眼腐烂果实中分离到的细菌和真菌目前在龙眼采后病害中未见相关报道,对于其致病性检测还有待于进一步的回接验证,以明确导致采后龙眼果实腐烂的主导微生物。     

Rye doubled haploids as a research and breeding tool – a practical point of view

Doubled haploid (DH) plants were produced using anther culture from out‐crossing rye, including breeders’ lines, cultivars and F₁ plants with DH parents, to examine the feasibility of using the DH technique for breeding and specifically for developing mapping populations. Only 10–36% of green regenerants produced via anther culture were suitable for research or breeding purposes because of low survival rate or low fertility. Spontaneously arising DH regenerants were more often fertile compared with the colchicine‐treated ones. The fertility of spontaneous DHs varied from sterile to half that found in a normal rye population, which has implications for the design of a crossing scheme and subsequent anther culture. In the reciprocal crosses within one DH population, fertility was the lowest observed, probably because of self‐incompatibility factors, whereas in the DH crosses with normal heterozygous cultivars fertility was the highest. Two mapping populations using DHs were established, the first for out‐crossing rye it would seem. These populations will be used for mapping two important traits, the semi‐dwarf growth habit and preharvest sprouting resistance in rye.     

在植株不同水平距离处垂直断根对夏玉米产量形成和籽粒库容特性的影响

以高产夏玉米品种郑单958(浅根型,ZD)和登海661(深根型,DH)为材料,于大喇叭口期(V12)分别在距离植株两侧10 cm、20 cm、30 cm处垂直断根,断根深度60 cm,以不断根处理为对照,共设计8个处理(ZDCK、ZD10、ZD20、ZD30;DHCK、DH10、DH20、DH30),研究了玉米植株两侧不同水平距离处根系对夏玉米籽粒灌浆及产量形成的调控作用。结果表明,ZD10、ZD20与DH10、DH20分别切断24.81%、11.69%与16.82%、7.52%的根系;ZD30与DH30各指标与CK间无显著差异,ZD10、ZD20产量分别下降13.09%、9.10%,显著大于DH10、DH20的产量降幅(9.81%、4.64%),其产量下降原因主要是断根后两品种的穗粒数与千粒重降低,其中ZD20、ZD10穗粒数与千粒重较ZDCK分别下降4.90%、5.60%和4.37%、7.88%,DH20、DH10穗粒数与千粒重较DHCK分别下降3.38%、5.15%和1.15%、4.97%;断根后两品种实际库容与结实率均有不同程度下降;到达最大灌浆速率时的天数(T_(max))、灌浆速率最大时的生长量(W_(max))、最大灌浆速率(G_(max))、籽粒灌浆活跃期(P)、灌浆速率也均下降,断根处理对浅根型品种郑单958的影响更为显著。     

Segregation of genetic markers among wheat doubled haploid lines derived from wheat x maize crosses

Summary Wheat doubled haploid (DH) lines were produced from the F₁ hybrid, Fukudo-komugi x Oligo Culm, through intergeneric crosses between wheat and maize. F₂ plants and 203 DH lines were analyzed for the segregation of the eight genetic markers, namely, grain proteins, grain esterases, GA-insensitivity and glume traits. The segregation in the F₂ plants fitted to the expected ratios. No deviation was observed among the DH lines, either, except for the glume pubescence. The result indicates the absence of correlation between the markers investigated and the efficiency of embryo formation in the DH lines.     

高密度条件下行距配置对春玉米光合特性及产量的影响

为探明在9.0万株/hm2高密度种植条件下旱地春玉米最佳的株行距配置,采用随机区组设计,12个处理(DH1.等行距40.0 cm,DH2.等行距50.0 cm,DH3.等行距60.0 cm,DH4.等行距70.0 cm,DH5.等行距80.0 cm,DH6.等行距90.0 cm,KH1.宽窄行(53.3 cm+26.7 cm),KH2.宽窄行(66.7 cm+33.3 cm),KH3.宽窄行(80.0 cm+40.0 cm),KH4.宽窄行(93.3 cm+46.7 cm),KH5.宽窄行(106.7 cm+53.3 cm),KH6.宽窄行(120.0 cm+60.0 cm))。3次重复,研究了行距配置对郑单958在旱地春播情况下产量和群体光合特性的影响。结果表明,随着行距的缩小,穗位叶SPAD值、穗位叶净光合速率、叶面积指数(LAI)、PAR截获率、单株干物质积累量、籽粒产量均提高;其中,50.0 cm+50.0 cm和66.7 cm+33.3 cm配置2年平均籽粒产量较90.0 cm+90.0 cm和120.0 cm+60.0 cm配置的平均产量提高24.3%,除2013年DH6处理产量显著高于KH6处理外,其余等行距处理和宽窄行处理之间产量无显著性差异。高密度条件下山西省春玉米最佳行距配置为50.0 cm+50.0 cm和66.7 cm+33.3 cm。