Improvement of Sclerotinia resistance of a Polima CMS restorer line of rapeseed via phenotypic selection, marker-assisted background selection and microspore culture

Sclerotinia stem rot caused by Sclerotinia sclerotiorum is one of the most severe diseases of rapeseed in China and other major growing regions. The objective of this study was to improve the S. sclerotiorum resistance of 'Hui5200', an elite 'Polima' CMS restorer line, by introgression and fixation of resistance alleles from the partially resistant cultivar 'NingRS-1' via phenotypic selection (PS), marker-assisted background selection (MAB) and microspore culture. A progeny designated as 'RSH' with greatly improved Sclerotinia resistance and a similar genetic background as 'Hui5200' was obtained by two backcrosses and one selfing. From a selected elite progeny line (named as 7-5) double haploid (DH) lines were developed. By three cycles of PS considering economic traits and genetic distance analysis, four resistant DH restorer lines with elite economic traits were finally selected. The obtained resistant restorer lines have been used to produce commercial F₁ hybrids. The results indicated that backcrossing plus PS and MAB is effective and suitable for improving resistance of rapeseed to S. sclerotiorum .     

RAPD markers linked to a clubroot-resistance locus in Brassica rapa L.

Linkage of random amplified polymorphic DNA (RAPD) markers with resistance genes to clubroot (Plasmodiophora brassicae Wor.) in Brassica rapa L. was studied in a doubled haploid (DH population obtained by microspore culture. Thirty-six DH lines were obtained from F₁ plants from a cross between susceptible ‘Homei P09’ and resistant ‘Siloga S2’ plants. ‘Homei P09’ was a DH line obtained by microspore culture of the Chinese cabbage variety ‘Homei’, which is highly responsive in microspore culture. The resistant line ‘Siloga S2’ was obtained by two rounds of selfing of the fodder turnip ‘Siloga’. Three RAPD markers, RA12-75A, WE22B and WE49B, were found to be linked to a clubroot-resistance locus. These three markers were linked in the DH lines and an F₂ population and should be useful for marker-assisted selection in breeding programs. This revised version was published online in July 2006 with corrections to the Cover Date.     

Field Performance of Androgenetic Doubled Haploid Spring Wheat Lines in Comparison with Lines Selected by the Pedigree System

A total of 316 doubled haploid lines (DH) of spring wheat were compared with 621 lines selected in a pedigree system (PS) under field conditions in a breeding nursery. The lines originated from 21 crosses and the samples tested represent mean values for variables comprising the main breeding goals such as disease resistance, baking quality, and agronomic traits. In general, the DH lines were later in days to heading and shorter than PS lines of different homozygosity levels. Except for leaf rust (Puccinia recondite), the DHs were more resistant to the artificially infected diseases such as powdery mildew (Erysiphe graminis), stripe rust (Puccinia striiformis), and septoria nodorum blotch (Septoria nodorum). These differences, which were relatively small for practical selection purposes, were probably due to easier and more precise disease assessment of the homozygous DH lines. For quality characters, the DHs had a higher protein level, while the rest of the parameters were similar for both origins. The data analyzed suggest so far that androgentic doubled haploids in spring wheat are very similar to lines selected in a pedigree system in respect to all the agronomic characters tested. However, the DH lines were produced in a much shorter period of time. It is suggested that androgenetic doubled haploids be produced from F₁ hybrids and that the well-established bulk method should continue to allow selection bulk method should continue to allow selection for rare recombinants as soon as homozygosity is reached.     

大白菜干烧心病抗性QTL定位研究

对大白菜干烧心病抗性不同的材料进行研究,明确其抗性遗传规律,并完成抗性基因的QTL定位分析,为分子标记辅助选择(MAS)育种与抗病机理的研究提供理论依据。采用大白菜干烧心病抗性显著不同的青麻叶类型高代自交系黑227(抗干烧心病)和包头型高代自交系B120(感干烧心病)作为材料,将得到的杂种F1进行小孢子培养得到DH群体,将亲本和F1及DH群体种植于日光温室,根据田间干烧心病发病程度进行分级,进而得出病情指数,并结合已构建的大白菜分子遗传图谱,利用Map QTL 5.0软件对大白菜干烧心病抗性基因进行定位。结果表明,试材所含有的大白菜干烧心病抗性基因符合数量性状遗传的特点,共检测到2个与大白菜干烧心病抗性基因连锁的InDel分子标记Br ID10343和Br ID10349,这2个标记均位于Chr.7,其间的遗传距离为1.031 c M,遗传贡献率均达到40%以上。InDel分子标记Br ID10343和Br ID10349与大白菜干烧心病抗性基因紧密连锁,结果为抗性基因主效QTL的精细定位及MAS抗病育种奠定了良好基础。     

水稻抗稻瘟病Pigm(t)基因的分子标记辅助选择与利用

旨在快速改良武运粳29196的稻瘟病抗性。采用定向回交育种策略,运用分子标记辅助选择技术和花药培养技术,快速改良武运粳29196的稻瘟病抗性。以籼稻品种谷梅4号为稻瘟病抗性基因Pigm(t)的供体,以高产易感稻瘟病的品系武运粳29196为受体,在连续回交和自交过程中,利用与Pigm(t)紧密连锁的In Dels标记S95477和S29742进行分子标记辅助选择。在BC2F1世代,进行花药培养,获得185个双单倍体群体(DH群体),从中筛选出82个含有Pigm(t)基因的改良株系。在经过对农艺性状、稻瘟病抗性的系统鉴定与稻米品质性状测定后,发现改良株系DH036和DH158的综合性状与武运粳29196已十分相近,且稻米品质有所提升,保持了武运粳29196丰产性的同时,稻瘟病抗性有了明显的提高。利用定向回交、花药培养和分子标记辅助选择相结合的技术体系,可明显提高稻瘟病抗性改良的预见性和准确性,缩短育种年限、加快育种进程。新育成的武运粳29196抗性改良系,为稻瘟病抗性育种提供了重要的遗传资源。     

Inheritance of resistance to flax wilt (Fusarium oxysporum f.sp. lini Schlecht) in a doubled haploid population of Linum usitatissimum L.

The inheritance of resistance to fusarium wilt (Fusarium oxysporum f.sp. lini) was investigated in Linum usitatissimum as a first step towards gaining an understanding of the molecular genetics of the disease and developing a procedure for marker-assisted selection. A recombinant doubled haploid (DH) population was derived from the haploid component of polyembryonic F₂ seeds originating from a cross between a wilt resistant, twinning Linola™ Linola is a registered trademark of CSIRO line CRZY8/RA91 and the wilt susceptible Australian flax cultivar Glenelg. The segregation of resistance was studied in 143 DH lines under glasshouse and field conditions. Most of the phenotypic variation was attributable to the segregation of two independent genes with additive effects. Minor resistance genes may have also contributed by modifying the resistance response. A glasshouse screening method of DH lines proved a reliable indicator of field resistance to fusarium wilt. This revised version was published online in July 2006 with corrections to the Cover Date.     

玉米自交系“南99”的选育及配合力分析

针对广西玉米育种材料遗传基础狭窄这一关键问题,此研究采用S1改良法群体内轮回选择与自交系谱选择相结合的育种方法,选育出"南99"自交系;经配合力测定和性状鉴定表明,"南99"是一个一般配合力和特殊配合力都高、综合农艺性状好、抗病虫性强、适应性广、自身产量高的自交系,用它做母本分别选育成南校18号、南校968、南校9665和南校201等四个优良玉米单交种并通过广西品种审定。其有较高的利用价值和广泛的应用前景。     

Development of novel clubroot resistant rapeseed lines (Brassica napus L.) effective against Japanese field isolates by marker assisted selection

Clubroot is an important disease infectible to cruciferous plants and a major threat to rapeseed production in Japan. However, no clubroot resistant rapeseed cultivars have been released. We surveyed pathotype variation of six isolates collected from rapeseed fields and found they were classified as pathotype groups 2 and 4 using Japanese F₁ Chinese cabbage cultivars. We produced the resynthesized clubroot resistant Brassica napus harboring two resistant loci, Crr1 and Crr2, by interspecific crossing and developed resistant rapeseed lines for southern and northern regions by marker-assisted selection and backcrossing. We improved the DNA marker for erucic acid content to remove linkage drag between Crr1 and high erucic acid content and successfully selected lines with clubroot resistance and zero erucic acid for northern regions. A novel line, ‘Tohoku No. 106’, suitable for southern regions showed stable resistance against all six isolates and high performance in infested fields. We conclude that Crr1 and Crr2 are important genes for CR rapeseed breeding and marker-assisted selection is effective in improving clubroot resistance.     

节瓜抗镰刀菌酸突变体的筛选和特性研究

枯萎病是节瓜的主要病害之一。为了创新节瓜抗枯萎病资源,利用枯萎病毒素镰刀菌酸（FA）为胁迫剂,以节瓜不定芽为筛选材料,进行了节瓜抗镰刀菌酸变异体离体筛选研究。试验结果表明,FA胁迫的适宜浓度为60~80mg•L-1。筛选出的抗性细胞系继代培养后仍保持对FA抗性,其再生植株的R1、R3代株系对节瓜枯萎病的抗性可以达到中抗至抗病级;与供体材料相比,再生系后代株系除枯萎病抗性增强外,其他主要农艺性状没有明显变化。表明利用抗FA细胞系离体筛选技术改良和创新节瓜抗枯萎病材料是可行的。     

Effect of thidiazuron on microspore embryogenesis and plantlet regeneration in Chinese flowering cabbage (Brassica rapa. var. parachinenis)

Traditional breeding methods require more than 6 years to obtain homozygous inbred lines, while isolated microspore culture (IMC) is an effective way to cultivate double haploid homozygous lines in only 2 years. However, low embryogenesis induction frequency in Chinese flowering cabbage remains a key obstacle to the practical application of this technique. Thidiazuron was added at different concentrations to NLN‐13 medium to estimate its effects on microspore embryogenesis and plantlet regeneration. Results showed that three genotypes responded positively. Optimum thidiazuron concentrations produced embryo yields of up to 14.67 embryos per bud and increased microspore embryogenesis frequency with up to 100% survival. Plantlet regeneration rates were up to 81.67%, and the treatment groups showed lower callus formation. We obtained up to 552 diploid plants from the tested genotypes, and the percentage of doubled haploid at different TDZ concentrations showed slight differences, and doubled haploid rates in the three genotypes were above 70%. They showed a high uniformity and can be directly used for hybrid breeding. This method accelerates microspore application in Chinese flowering cabbage hybrid breeding.     

Combining ability for beet armyworm, Spodoptera exigua, resistance and horticultural traits of selected Lycopersicon pennellii-derived inbred backcross lines of tomato

Ten selected inbred backcross lines (IBL), from a Lycopersicon esculentum cv.‘Peto 84’×Lycopersicon pennellii IBL population, with resistance to beet armyworm (BAW), Spodoptera exigua, higher fruit mass and fruit yield, were crossed with eight elite cultivated L. esculentum inbred lines in a Design II mating design. Three elite inbreds were also crossed to ‘Peto 84′, the IBL recurrent parent, as a control for combining ability. Field plots of all resulting F₁ progenies and control cultivars were inoculated with BAW eggs and evaluated for resistance to BAW, fruit mass, fruit yield, vine size and maturity at three field locations. Reductions in fruit damage by BAW were found in four of the 10 IBL F₁ progenies. Significant male and female general combining ability (GCA) estimates for BAW resistance were observed, but significant specific combining ability for BAW resistance was not detected. The fruit mass of F₁ hybrids was significantly lower than large-fruited controls, but was not significantly different from elite inbred by ‘Peto 95’F₁ hybrids. Selection based on inbred performance identified IBL with positive GCA for BAW resistance and yields in Design II hybrids. BAW resistance in the 09 selected IBL and IBL- derived F₁ progeny was associated with two undesirable traits, later maturity and larger vine size. Index selection of IBL was more effective at identifying IBL with positive GCA for fruit mass and fruit yield than GCA for BAW resistance.     

油菜小孢子培养技术体系研究

小孢子培养在油菜的基础研究和应用研究中均具有十分重要的意义。自1982年Lichter首次在甘蓝型油菜中进行小孢子培养获得成功以来,国内外在油菜小孢子培养技术方面已取得大量研究成果,包括油菜小孢子胚状体发生的影响因素,小孢子植株的再生、成苗、大田移栽、染色体加倍等,近年来又对一些关键技术环节加以了改进,笔者在对这些研究成果进行总结的基础上针对中国国情建立了大田条件下油菜高效小孢子培养技术体系。用该体系对甘蓝型油菜和新疆野生油菜的体细胞杂种后代进行小孢子培养的出胚率达到300枚/皿以上,采用小孢子苗直接移栽大田技术,成活率达到89.0%。此外还成功构建了含127个DH系的黄籽油菜DH群体及含115个DH系的粒重分离群体。     

分子标记辅助选育水稻抗白叶枯病和稻瘟病多基因聚合恢复系

利用分子标记辅助选择和田间鉴定选择相结合的方法, 将三黄占2号的抗稻瘟病主基因Pi-GD-1(t)、Pi-GD-2(t)和主效QTL GLP8-6(t) (分别简称G1、G2和G8)及抗白叶枯病基因Xa23导入到明恢86、蜀恢527和浙恢7954等3个骨干中籼恢复系, 通过复交进行基因聚合, 获得5个带有抗稻瘟病兼抗白叶枯病的双基因或多基因聚合系明恢86-G1-G2-Xa23、蜀恢527-G2-Xa23、明浙-G2-G8-Xa23-1、明浙-G2-G8-Xa23-2和明浙-G1-G2-G8-Xa23。以上5个抗病基因聚合改良系对稻瘟病的抗谱与抗源品种相仿或更宽, 改良系和与不育系II-32A配制的测交种对白叶枯病菌的抗谱与供体亲本IRBB23一致, 测交种在不接种白叶枯病菌条件下的产量和结实率与原来的恢复系及相应杂交种相仿, 但在接种条件下带有Xa23基因的恢复系及测交种的结实率、千粒重和产量明显优于原来的恢复系及相应杂交种。研究表明, 抗稻瘟病基因和抗白叶枯基因Xa23在不同恢复系背景下的抗性表达完全, 对恢复系稻瘟病以及白叶枯病改良的效果明显。对分子标记复交改良恢复系的抗病性进行了讨论。     

中国玉米抗病育种的若干问题及其解决途径

我国玉米生产中的主要病害有大斑病、小斑病、丝黑穗病、茎腐病和矮花叶病。研究结果表明:玉米对上述病害的抗性,主要表现为受多基因控制的水平抗性。不同玉米材料,抗病性有明显差别。F_1的抗病性,倾向抗病亲本或比亲本的平均病级低。实践证明,采用轮回选择法和利用多细胞质杂交种是行之有效的玉米抗病育种途径。     

培育水稻恢复系抗稻褐飞虱基因导入系和聚合系

稻褐飞虱是水稻的主要虫害之一,培育优良的抗性基因聚合系对于防治稻褐飞虱具有重要的意义.本研究通过回交、分子标记辅助选择和接虫鉴定三者相结合的办法,将抗稻褐飞虱基因Bph3和Bph24(t)分别导入主栽杂交水稻恢复系广恢998、9311、R15、明恢63、R29中,最终获得遗传稳定的Bph3导入系32份和Bph24(t)...     

Inheritance of resistance to Fusarium head blight in the wheat lines 'CJ 9306' and 'CJ 9403'

Fusarium head blight (FHB or scab) caused by Fusarium graminearum is a worldwide serious disease in wheat. Exploitation and genetic studies of elite resistance sources can speed up the development of resistant cultivars. To characterize the inheritance of host plant resistance in two new lines, ‘CJ 9306’ and ‘CJ 9403’, developed from a recurrent selection programme in China, six generations P₁, P₂, F₁, F₂, B₁ and B₂ of four crosses and 137 F_{6 : 7} recombinant inbred lines (RILs) from one cross were evaluated in the greenhouse for scab resistance using single‐floret inoculation. The data of area under disease progress curve (AUDPC) in F₂, backcross (BC) and RIL populations exhibited mono‐modal distributions without clear‐cut demarcations and skewing towards resistance. An additive–dominance model was well‐fitted, additive effects played a predominating role, and dominance effects were also significant. Continuous distributions with two major peaks and one minor peak for the number or percentage of scabby spikelets (NSS or PSS) in segregating populations implied the existence of major genes or quantitative trait loci (QTL) for resistance. The estimates of broad‐sense and narrow‐sense heritabilities based on the six‐generation experiment were 56–76% and 26–67% respectively. The estimates of broad‐sense heritabilities based on anova with RILs were 89–90%. These two improved lines with excellent scab resistance and good agronomic traits are of interest for wheat breeding and production.     

The major quantitative trait locus for mungbean yellow mosaic Indian virus resistance is tightly linked in repulsion phase to the major bruchid resistance locus in a cross between mungbean [Vigna radiata (L.) Wilczek] and its wild relative Vigna radiata ssp. sublobata

Mungbean yellow mosaic Indian virus (MYMIV) and bruchid infestation are severe production constraints of mungbean in South Asia, a major global mungbean production area. Marker-assisted selection for resistance against these disorders while maintaining or even improving agronomic traits is an important step toward breeding elite mungbean varieties. This study employed recombinant inbred lines (F12) derived from a cross between MYMIV-tolerant Vigna radiata NM92 and bruchid-resistant V. radiata ssp. sublobata TC1966 to identify chromosomal locations associated with disease and insect pest resistance and seed traits. A linkage map comprising 11 linkage groups was constructed with random amplified polymorphic DNA (RAPD), sequence characterized amplified regions (SCAR), cleaved amplified polymorphic DNA (CAP), amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. Quantitative trait loci (QTLs) for MYMIV and bruchid resistance, 100 seed weight and seed germination rate were identified. Three major QTLs for MYMIV and one major bruchid resistance locus were mapped on LG 9. The resistance alleles were contributed by the MYMIV tolerant parent NM92 and the bruchid resistant parent TC1966 respectively. One of the MYMIV QTLs was tightly linked in repulsion phase to the bruchid resistance locus. In addition, three minor QTLs for MYMIV resistance were found, where the resistance alleles were contributed by TC1966. Lines combining MYMV resistance alleles from both parents have greater resistance to MYMIV than the tolerant parent. Two minor bruchid resistance QTLs were identified in TC1966. Furthermore, three QTLs each for 100 seed weight and germination rate were detected. The markers defining the QTLs identified in this study will be useful in marker-assisted breeding of improved mungbean varieties in the future.     

Improved doubled haploid production protocol for Brassica napus using microspore colchicine treatment in vitro and ploidy determination by flow cytometry

A rapid and efficient microspore culture protocol was applied to produce homozygous progeny of crosses between low erucic canola and high erucic resynthesized rapeseed (Brassica napus L.). Microspores of Canadian cultivars ‘Excel’ and ‘Profit’ as well as three F₁ hybrids with the resynthetic line ‘RS239’ were treated with colchicine immediately after isolation. Flow cytometry was applied for early identification of doubled haploid (DH) regenerants. The diploidization rate was subsequently verified by scoring flower morphology. In vitro colchicine treatment had a positive effect on induced diploidization, and was associated with the frequency of preliminary spontaneous diploidization which was, however, determined by the genotype. In addition, the effects of colchicine treatment on embryoid formation and regeneration have been evaluated. The method presented is feasible for commercial large‐scale production of DHs in rapeseed as the genotype‐specific diploidization can be efficiently balanced by in vitro colchicine treatment. In addition, the use of flow cytometry immediately after in vitro culture allows efficient selection for DHs, thus saving labour and cost and in the laboratory and subsequent greenhouse phase.     

waxy基因功能标记开发及在糯玉米育种中的应用

为利用分子标记辅助选择加快糯玉米种质资源改良进程,以糯玉米自交系云539和普通玉米自交系昌7-2为材料,通过DNA序列测定和分析,探明糯玉米自交系云539 wx基因属于wx-D7突变类型。根据2个自交系基因组序列差异,开发出waxy基因功能标记FMD7,并通过了表型验证和通用性验证。利用开发的基因功能标记FMD7进行分子标记辅助选择,结合产量、株型和品质性状分析,获得携带wx基因的优良昌7-2改良系6个。利用本研究开发的基因功能标记FMD7进行辅助选择可加快回交转育进程,提高育种效率,为选育高产糯玉米杂交种提供材料基础,同时也为分子标记辅助选择单基因或少数主效基因控制的性状提供理论参考。     

Characterization of Chinese wheat germplasm for resistance to Fusarium head blight at CIMMYT, Mexico

Fusarium head blight (FHB) poses a challenge for wheat breeders worldwide; there are limited sources of resistance and the genetic basis for resistance is not well understood. In the mid-1980s, a shuttle breeding and germplasm exchange program launched between CIMMYT-Mexico and China, enabled the incorporation of FHB resistance from Chinese bread wheat germplasm into CIMMYT wheat. Most of the Chinese wheat materials conserved in the CIMMYT germplasm bank had not been fully characterized for FHB reaction under Mexican environments, until 2009, when 491 Chinese bread wheat lines were evaluated in a FHB screening nursery in Mexico, and 304 (61.9 %) showed FHB indices below 10 %. Subsequent testing occurred in 2010 for plant height (PH), days to heading (DH), and leaf rust response. In 2012, 140 elite lines with good agronomic types were further evaluated for field FHB reaction and deoxynivalenol (DON) accumulation. Most of the tested lines showed good resistance: 116 (82.9 %) entries displayed FHB indices lower than 10 %, while 89 (63.6 %) had DON contents lower than 1.0 ppm. Significant negative correlations were observed between FHB traits (FHB index, DON content, and Fusarium damaged kernels) and PH, DH, and anther extrusion. A subset of 102 elite entries was selected for haplotyping using markers linked to 10 well known FHB quantitative trait loci (QTL). 57 % of the lines possessed the same 2DL QTL marker alleles as Wuhan 1 or CJ 9306, and 26.5 % had the same 3BS QTL allele as Sumai 3. The remaining known QTL were of low frequency. These materials, especially those with none of the above tested resistance QTL (26.5 %), could be used in breeding programs as new resistance sources possessing novel genes for FHB resistance and DON tolerance.