Construction of a molecular functional map of rapeseed (Brassica napus L.) using differentially expressed genes between hybrid and its parents

Brassica napus L. is an important oilseed and fodder crop with significant heterosis for seed yield and other agronomic traits, but very little is known about the molecular basis of heterosis. As an initial step towards understanding the molecular events associated with this phenomenon, a molecular functional map of rapeseed was constructed using differentially expressed genes in hybrid identified by microarrays. Single-strand conformational polymorphism (SSCP) analysis was applied for genetic mapping in an F ₂ population of 184 individuals resulting from crossing ‘`SI-1300 × Eagle’'. A total of 162 markers including 154 loci corresponding to 98 differentially expressed genes assigned to 17 functional categories and 8 SSR markers were grouped into 21 linkage groups (LGs), covering a total map distance of 2267.3 cM. Subsequently, this map was aligned with Arabidopsis thaliana in silico. Comparative mapping shows that genes localized on each Arabidopsis chromosome have orthologs dispreading in different B. napus LGs. Similarly, a majority of LGs were made of homologous genes from different Arabidopsis chromosomes. In addition, a total of 25 syntenic regions were identified in B. napus, in most of which the gene order was not consistent between the two species, and each of the conserved regions in the A. thaliana genome was homologous to 1--5 distinct regions in the B. napus genome. These results indicate that it is not easy to exploit A. thaliana information for B. napus based on synteny.     

In silico-selection of Brassica rapa organelle genome-derived BACs using their end sequences and sequence level comparative analysis of the 124 kb mitochondrial genome sequences in the family Brassicaceae

We identified BAC clones which harbor DNAs derived from the B. rapa organelle genomes by in silico mapping of 80,292 B. rapa BAC end sequences on the Arabidopsis organelle genomes and subsequent insert size estimation and fingerprinting. A total of 1,048 putative chloroplast genome-derived BAC clones (2.6%) were identified. Fingerprinting and sequencing revealed that many of them represented the entire chloroplast genome (about 150 kb). Meanwhile, only 59 putative mitochondrial genome-derived BACs (0.15%) were identified and most of them showed rare agreement between the in silico map and fingerprinting. We sequenced BAC clone KBrB042G11 (42G11) and compared it to the mitochondrial genome of B. napus and A. thaliana which showed dynamic rearrangement events. The order of 33 orthologous genes was collinear between the 42G11 BAC and its counterpart in B. napus. Five distinctive rearrangements and two InDels were identified between these two closely related species and the rearrangements were related to the occurrence of small tandem repeat sequences. Sequences of the 33 orthologous genes in the homoeologous regions of B. napus and B. rapa were almost 100% identical. Gene orders showed no colinearity between Arabidopsis and Brassica even though 31 orthologous genes shared high sequence similarity with p-values over 1E-32. FISH analysis using the identified BAC revealed a large chloroplast genome insertion in the pericentromeric region of chromosome (chr.) 4 of B. rapa.     

海岛棉A亚组和草棉及阿非利加棉单染色体鉴定

 以1套13个陆地棉A亚组染色体特异的BAC克隆为探针, 与海岛棉Pima90-53、红星草棉和阿非利加棉染色体进行荧光原位杂交。这些探针均在染色体上产生了特异信号,从而鉴别了这3个棉种基因组的单染色体。因此,这套BAC克隆也可以作为这3个基因组染色体的细胞遗传学标记。在此基础上,对3个棉种的A（亚）组染色体物理序号进行了命名。根据这些分子标记和连锁群的关系,染色体物理序号和遗传连锁图谱间可实现相互转化。这将有助于研究棉属不同棉种间的起源演化关系,以及新的遗传标记的染色体物理定位和构建染色体物理图谱等。     

Persistent C genome chromosome regions identified by SSR analysis in backcross progenies between Brassica juncea and B. napus

Given that feral transgenic canola (Brassica napus) from spilled seeds has been found outside of farmer’s fields and that B. juncea is distributed worldwide, it is possible that introgression to B. juncea from B. napus has occurred. To investigate such introgression, we characterized the persistence of B. napus C genome chromosome (C-chromosome) regions in backcross progenies by B. napus C-chromosome specific simple sequence repeat (SSR) markers. We produced backcross progenies from B. juncea and F₁ hybrid of B. juncea × B. napus to evaluate persistence of C-chromosome region, and screened 83 markers from a set of reported C-chromosome specific SSR markers. Eighty-five percent of the SSR markers were deleted in the BC₁ obtained from B. juncea × F₁ hybrid, and this BC₁ exhibited a plant type like that of B. juncea. Most markers were deleted in BC₂ and BC₃ plants, with only two markers persisting in the BC₃. These results indicate a small possibility of persistence of C-chromosome regions in our backcross progenies. Knowledge about the persistence of B. napus C-chromosome regions in backcross progenies may contribute to shed light on gene introgression.     

Sexual transfer of Arabidopsis DNA to Brassica napus

Out of 400 reciprocal crosses made between Brassica napus and Arabidopsis thaliana, 200 maternal B. napus flowers produced 31 seeds and 10 seeds developed when A. thaliana was used as the female parent. Of the latter, three seeds successfully germinated but did not develop into mature plants. In contrast, plants from all 32 seeds generated from B. napus were established. DNA from these 31 plants was analysed by 18 probes mapped to the A. thaliana genome, two selectable marker genes and one A. thaliana species-specific repetitive probe. It was found that one plant had identical restriction fragments to A. thaliana with two of the 21 probes used. The gene flow within Brassicaceae is discussed in light of this result.     

Inheritances and location of powdery mildew resistance gene in melon Edisto47

Powdery mildew caused by Podosphaera xanthii is a major disease in melon. Here we report two Px race 1 strains named Px1A and Px1B in Xinjiang, which have different pathogenicities. The more pathogenic Px1B made some powdery mildew resistant genes on linkage group V (LGV) lose their resistant traits. The inheritances of resistance to Px1A and Px1B in melon Edisto47 were studied using a BC₁ population derived from a cross between the resistant genotype Edisto47 and the susceptible cultivar Queen. The resistance/susceptibility segregation ratios observed in the Px1A-inoculated BC₁ population and the loci of polymorphic markers indicated that resistance to Px1A was controlled by two dominant genes. Quantitative trait locus analysis identified two loci mapped on LGII and LGV, respectively, for powdery mildew resistance. However, for resistance to Px1B, Edisto47 was found to bear one dominant gene. A genetic linkage map was constructed using the Px1B-inoculated BC₁ population to map the resistant gene. Comparative genomic analyses revealed that the linkage map of Pm-Edisto47-1 was collinear with the corresponding genomic region of the melon chromosome 2. Genetic analysis showed that Pm-Edisto47-1 was located between simple sequence repeat (SSR) markers CMGA36 and SSR252089, at a genetic distance of 2.1 cM to both markers. Synteny analysis showed that two genes named MELO3C015353 and MELO3C015354 were predicted as candidates for Edisto47-1 in this region.     

Molecular mapping of a stripe rust resistance gene in wheat cultivar Wuhan 2

Stripe rust (or yellow rust), caused by Puccinia striiformis f. sp. tritici, is one of the most destructive diseases of wheat worldwide. Growing resistant cultivars is the best approach to control the disease. To identify and map genes for stripe rust resistance in wheat cultivar ‘Wuhan 2', an F₂ population was developed from a cross between the cultivar and susceptible cultivar Mingxian 169. The parents, 179 F₂ plants and their derived F_2:3 lines were evaluated for responses to Chinese races CYR30 and CYR31 of the pathogen in a greenhouse. A recessive gene for resistance was identified. DNA bulked segregant analysis was applied and resistance gene analog polymorphism (RGAP) and simple sequence repeat (SSR) techniques were used to identify molecular markers linked to the resistance gene. A genetic map consisting of five RGAP and six SSR markers was constructed. The recessive gene, designated Yrwh2, was located on the short arm of chromosome 3B and flanked by SSR markers Xwmc540 and Xgwm566 at 5.9 and 10.0 cM, respectively. The chromosomal location of the resistance gene and its close marker suggest that the locus is different from previously reported stripe rust resistance genes Yr30, QYr.ucw-3BS, Yrns-B1, YrRub and QYrex.wgp-3BL previously mapped to chromosome 3B. Yrwh2 and its closely linked markers are potentially useful for developing stripe rust resistance wheat cultivars if used in combination with other genes.     

Genetic mapping of bph20(t) and bph21(t) loci conferring brown planthopper resistance to Nilaparvata lugens St?l in rice (Oryza sativa L.)

Brown planthopper(BPH) is one of the most serious and destructive insect pests of rice in most rice growing regions of the world. In this study, two major resistance genes against BPH have been identified in an Oryza rufipogon (Griff.) introgression rice line, RBPH54. Inheritance of the BPH resistance in RBPH54 was studied by screening the resistance in parents, F1, F2 and BC1 generations against BPH biotype 2. A population of BC3F2 lines was developed and SSR markers were employed for the gene mapping, and new markers were designed for fine mapping of the resistance genes, while sequence information of BAC/PAC clones was used to construct physical maps of the genes. The results showed that the BPH resistance in RBPH54 was governed by recessive alleles at two loci, tentatively designated as bph20(t) and bph21(t). The locus bph20(t) was fine mapped to the short arm of chromosome 6 about 2.7 cM to the upper marker RM435 and 2.5 cM to lower marker RM540 and in a 2.5 cM region flanked by two new SSR markers BYL7 and BYL8 which were developed in the present study. The other BPH resistance locus bph21(t) was initially mapped to a region 7.9 cM to upper marker RM222 and 4.0 cM to lower marker RM244 on the short arm of chromosome 10. For physical mapping, the bph20(t)-linked markers were landed on BAC/PAC clones of the reference cv., Nipponbare, released by the International Rice Genome Sequencing Project. The bph20(t) locus was physically defined to an interval of about 75 kb with clone P0514G1. Identification and location of these two genes in the present study have diversified the BPH resistance gene pool, which give benefit to the development of resistant rice cultivars, and the linkage PCR-based SSR markers for the bph20(t) and bph21(t) genes would help realize the application of the genes in rice breeding through marker-assisted selection.     

Analysis of QTL for seed oil content in <Emphasis Type="Italic">Brassica napus</Emphasis> by association mapping and QTL mapping

Increasing seed oil content is one of the most important breeding targets for rapeseed (Brassica napus). In this study, we combined quantitative trait loci (QTL) mapping and marker-trait association analysis to dissect the genetic basis of seed oil content in rapeseed. A doubled haploid (DH) population with 261 lines was grown in two highly contrasting macro-environments, Germany with winter ecotype environment and China with semi-winter ecotype environment, to explore the effect of environment effect of on seed oil content. Notable macro-environment effect was found for seed oil content. 19 QTL for seed oil content were identified across the two macro-environments. For association analysis, a total of 142 rapeseed breeding lines with diverse oil contents were grow in China macro-environment. We identified 23 simple sequence repeat (SSR) markers that were significantly associated with the seed oil content. Comparative analysis revealed that five QTL identified in the DH population, located on chromosomes A03, A09, A10 and C09, were co-localized with 11 significantly associated SSR markers that were identified from the association mapping population. Of which, the QTL on chromosome A10 was found to be homeologous with the QTL on chromosome C09 by aligning QTL confidence intervals with the reference genomes B. napus. Those QTL associated with specific macro-environments provides valuable insight into the genetic regulation of seed oil content and will facilitate marker-assisted breeding of B. napus.     

Genetic analysis of seed mineral accumulation affected by phosphorus deprivation in Brassica napus

The mineral content of plant seeds depends on both environmental and genetic factors. The aim of this study was to detect quantitative trait loci (QTLs), and their candidate genes, for the accumulation of phosphorus (P), calcium (Ca), magnesium (Mg), zinc (Zn), copper (Cu), iron (Fe), and manganese (Mn) in seeds of Brassica napus under normal and low P conditions using an F ₁₀ recombinant inbred line (RIL) population. Two-year field trials were conducted to investigate seed mineral accumulation. The results showed a significant decrease in most of the minerals in the BE RIL population, as well as in two parental lines, when grown in a low P environment compared to a normal P environment. In total, 60 putative QTLs were identified, 33 of which overlapped with each other in nine genomic regions in seven linkage groups. Twenty-one of the 60 significant QTLs co-located with eight seed weight QTLs, and only five overlapped with three seed yield QTLs. Moreover, only six QTLs for the same minerals were identified both in normal and low P levels. By comparative mapping of Arabidopsis and B. napus, 148 orthologs of 97 genes involved in the homeostasis of the seven minerals in Arabidopsis were associated with 47 QTLs corresponding to 24 chromosomal regions. These results offer insight into the genetic basis of mineral accumulation across different P conditions in seeds of B. napus and allow the potential utilization of QTLs in biofortification.     

Development of tomato SSR markers from anchored BAC clones of chromosome 12 and their application for genetic diversity analysis and linkage mapping

In this study, we developed a total of 37 simple sequence repeat (SSR) markers from 11 bacterial artificial chromosome (BAC) clone sequences anchored on chromosome 12 of tomato available at Solanaceae Genomics Network. These SSR markers could group a set of 16 tomato genotypes comprising of Solanum lycopersicum, S. pimpinellifolium, S. habrochaites, and S. pennellii unambiguously according to their known species status. Clear subgroups of genotypes within S. lycopersicum were also observed. A subset of 16 SSR markers representing the 11 BAC clones was used for developing genetic linkage maps of three interspecific F₂ populations produced from the crosses involving a common S. lycopersicum parent (CLN2498E) with S. pennellii (LA1940), S. habrochaites (LA407) and S. pimpinellifolium (LA1579). The length of the genetic linkage maps were 112.5 cM, 109.3 cM and 114.1 cM, respectively. Finally, an integrated genetic linkage map spanning a total length of 118.7 cM was developed. The reported SSR markers are uniformly distributed on chromosome 12 and would be useful for genetic diversity and mapping studies in tomato.     

Assessing and broadening genetic diversity of a rapeseed germplasm collection

Assessing the level of genetic diversity within a germplasm collection contributes to evaluating the potential for its utilization as a gene pool to improve the performance of cultivars. In this study, 45 high-quality simple sequence repeat (SSR) markers were screened and used to estimate the genetic base of a world-wide collection of 248 rapeseed (Brassica napus) inbred lines. For the whole collection, the genetic diversity of A genome was higher than that of C genome. The genetic diversity of C genome for the semi-winter type was the lowest among the different germplasm types. Because B. oleracea is usually used to broaden the genetic diversity of C genome in rapeseed, we evaluated the potential of 25 wild B. oleracea lines. More allelic variations and a higher genetic diversity were observed in B. oleracea than in rapeseed. One B. oleracea line and one oilseed B. rapa line were used to generate a resynthesized Brassica napus line, which was then crossed with six semi-winter rapeseed cultivars to produce 7 F₁ hybrids. Not only the allele introgression but also mutations were observed in the hybrids, resulting in significant improvement of the genetic base.     

Identification and multiple comparisons of QTL and epistatic interaction conferring high yield under boron and phosphorus deprivation in Brassica napus

Boron (B) and phosphorus (P) are two essential nutrients for plants. To unravel the genetic basis of B and P efficiency in Brassica napus, quantitative trait locus (QTL) and epistatic interaction analysis for yield and yield-related traits under contrast B and P conditions were performed using two mapping populations across various environments. Main effect QTLs were detected by QTLNetwork and QTL Icimapping (ICIM), and were compared with our previously reported main effect QTLs identified by QTLCartographer. Epistatic QTLs were identified by QTLNetwork, ICIM and Genotype matrix mapping (GMM), and multiple comparisons of main effect QTLs and epistatic QTLs were conducted. For the two mapping populations, 51 main effect QTLs were identified by QTLNetwork, 106 by ICIM. Among them, 35 main effect QTLs were simultaneously identified by three programs. Moreover, 578, 18 and 62 epistatic QTLs were identified by GMM, QTLNetwork and ICIM, respectively. Interestingly, a total of 235 epistatic QTLs identified by GMM were associated with 50 main effect QTLs identified by three programs. However, only nine epistatic QTLs identified by QTLNetwork and ICIM were involved in main effect QTLs. Twenty-two main effect QTLs in the BERIL population overlapped with 20 main effect QTLs for the same traits in the BQDH population, but no main effect QTLs were detected both under P and B stress environments, indicating the genetic differences in B and P homeostasis in B. napus. By in silico mapping, 29 candidate genes were located in the consensus QTL intervals. This study suggested the availability of dissecting genetic basis for complex traits under B/P deficiency by analyzing main effect QTLs and epistatic QTLs using multiple programs across different environments. The robust main effect QTLs and epistatic QTLs associated could be useful in breeding B and P efficient cultivars of B. napus.     

Identification of tightly linked SSR markers for flower type in carnation (Dianthus caryophyllus L.)

Single or double flower type is one of the most important breeding targets in carnation (Dianthus caryophyllus L.). We mapped the D ₈₅ locus, which controls flower type, to LG 85P_15–2 using a simple sequence repeat (SSR)-based genetic linkage map constructed using 91 F₂ progeny derived from a cross between line 85–11 (double flower) and ‘Pretty Favvare’ (single flower). A positional comparison using SSR markers as anchor loci revealed that the map positions of the D ₈₅ locus corresponded to the single locus controlling the single flower type derived from wild D. capitatus ssp. andrzejowskianus. We identified four co-segregating SSR markers on the D ₈₅ locus. Verification of the SSR markers in commercial cultivars revealed that two of the four SSR markers (CES0212 and CES1982) were tightly linked to the D ₈₅ locus, and amplified a 176-bp and 269-bp allele, respectively, which were common and unique to double flower cultivars. The map positions of the D ₈₅ locus and the tightly linked SSR markers will be useful for determining the genetic basis of flower type and for marker-assisted breeding of carnations.     

芥菜型油菜多室基因Bjmc2的精细定位

芥菜型多室油菜的产量比普通两室油菜更高,定位乃至克隆多室基因可为油菜遗传改良及解释多室角果形成机制创造条件。本研究通过验证JD11-2家系衍生群体仅在BjMc2位点上存在差异,可用于BjMc2的定位。采用AFLP结合BSA法分析BC₅和BC₆群体,筛选到1个与BjMc2连锁的AFLP标记并转化为SCAR标记SC1。基于该AFLP标记序列信息,利用白菜同源序列设计SSR引物和SCAR引物,获得11对SSR标记和1对SCAR标记。通过在芥菜型油菜BAC文库中的挑选,获得2个覆盖目标区域的单克隆,由此开发1个SSR标记。将获得的SCAR和SSR标记扫描BC₇群体,构建了两室性状基因BjMc2的遗传连锁图,两侧最近标记ZX17和BACsr96与目标基因之间的遗传距离分别为0.048 cM和0.340 cM,并定位到白菜A7 scaffold000019的946~1014 kb之间,约68 kb物理距离。     

Development and characterization of tomato SSR markers from genomic sequences of anchored BAC clones on chromosome 6

Simple sequence repeat motifs are abundant in plant genomes and are commonly used molecular markers in plant breeding. In tomato, currently available genetic maps possess a limited number of simple sequence repeat (SSR) markers that are not evenly distributed in the genome. This situation warrants the need for more SSRs in genomic regions lacking adequate markers. The objective of the study was to develop SSR markers pertaining to chromosome 6 from bacterial artificial chromosome (BAC) sequences available at Solanaceae Genomics Network. A total of 54 SSR primer pairs from 17 BAC clones on chromosome 6 were designed and validated. Polymorphism of these loci was evaluated in a panel of 16 genotypes comprising of Solanum lycopersicum and its wild relatives. Genetic diversity analysis based on these markers could distinguish genotypes at species level. Twenty-one SSR markers derived from 13 BAC clones were polymorphic between two closely related tomato accessions, West Virginia 700 and Hawaii 7996 and were mapped using a recombinant inbred line population derived from a cross between these two accessions. The markers were distributed throughout the chromosome spanning a total length of 117.6 cM following the order of the original BAC clones. A major QTL associated with resistance to bacterial wilt was mapped on chromosome 6 at similar location of the reported Bwr-6 locus. These chromosome 6-specific SSR markers developed in this study are useful tools for cultivar identification, genetic diversity analysis and genetic mapping in tomato.     

SCAR and CAPS markers flanking the Brassica oleracea L. Pp523 downy mildew resistance locus demarcate a genomic region syntenic to the top arm end of Arabidopsis thaliana L. chromosome 1

We recently mapped the Pp523 locus that includes a single, dominant gene conferring resistance to downy mildew expressed in adult plants to a 75.1 cm long linkage group on a genetic linkage map of Brassica oleracea L. More recently, we identified a new AFLP marker 2.8 cm downstream from the resistance gene. The five DNA markers within an 8.5 cm region encompassing the Pp523 gene were cloned and sequenced. Three of these markers were transformed into SCARs (sequence characterised amplified regions), however, two among them were monomorphic and were analysed as CAPS (cleaved amplified polymorphic sequence) markers among the mapping population. Searched against genomic databases, the five B. oleracea DNA-marker sequences matched Arabidopsis thaliana L. gene sequences that delimit a conserved syntenic region in the top arm end of chromosome 1 of this last species. Considering the close genetic relatedness between both species, the information on this specific genomic region in A. thaliana is particularly useful for the construction of a fine-scale map of the corresponding genomic region in B. oleracea. The identified SCAR and CAPS markers can be used for marker assisted selection (MAS) in breeding programs aimed at the introgression of the Pp523 resistance locus, allowing the reliable indirect identification of plants harbouring the resistance gene with a margin of error of approximately six in ten-thousand selected plants.     

Efficiency of PCR-RF-SSCP marker production in Brassica oleracea using Brassica EST sequences

Efficiencies of SCAR, CAPS and PCR-RF-SSCP marker production were investigated using two combinations of breeding lines in Brassica oleracea. Published EST sequences of B. oleracea, Brassica rapa, Brassica napus, and Arabidopsis thaliana and newly determined nucleotide sequences of anther cDNA clones from B. oleracea were used for designing primer pairs to amplify genes. The percentage of primer pairs yielding DNA amplification of a single gene was higher in primer pairs of B. oleracea (91%) than those of B. rapa (56%) and A. thaliana (17%). Single DNA fragments amplified by 9% of the primer pairs showed polymorphism as SCAR markers between a broccoli line and a Chinese kale line by agarose-gel electrophoresis. CAPS analysis showed different band patterns in 32% of the same-sized DNA fragments, and PCR-RF-SSCP analysis revealed DNA polymorphism in 52% of those showing no DNA polymorphism by CAPS. In total, 71% of the single DNA fragments were converted to DNA markers. The frequency of DNA polymorphism between parental lines of a cabbage F₁ hybrid was lower, 5% by SCAR and 12% by CAPS. However PCR-RF-SSCP analysis revealed DNA polymorphism in 21% of the DNA fragments showing no polymorphism by CAPS. These results suggest that PCR-RF-SSCP analysis enables highly efficient DNA marker production for mapping of genes in Brassica using progeny, even progeny of closely related parents. Analysis of selfed seeds of broccoli F₁ cultivars using PCR-RF-SSCP markers indicated that PCR-RF-SSCP analysis is also applicable to seed purity tests.     

Map- vs. homology-based cloning for the recessive gene ol-2 conferring resistance to tomato powdery mildew

The recessive gene ol-2 confers papilla-associated and race-non-specific resistance to tomato powdery mildew caused by Oidium neolycopersici. In order to facilitate marker assisted selection (MAS) in practical breeding programmes, we identified two simple sequence repeat (SSR) markers and one cleaved amplified polymorphic sequence (CAPS) marker which are linked to the resistance locus and co-dominantly inherited. Aiming to provide a base for ol-2 positional cloning, we used a large segregating F₂ population to merge these markers with all the ol-2 linked amplified fragment length polymorphism (AFLP^®) markers previously identified in an integrated genetic map. By screening a tomato bacterial artificial chromosome (BAC) library, we detected two BAC clones containing two expressed sequence tags (ESTs) homologous to the gene mlo, responsible for powdery mildew resistance in barley, as well as an ol-2-linked marker. Chromosomal mapping by Fluorescence in situ Hybridization (FISH) revealed major signals of the two BAC DNAs in the pericentromeric heterochromatin of the short arm of chromosome 4, in the same region where the ol-2 gene was previously mapped. The genetic and cytogenetic co-localisation between ol-2 and tomato mlo-homologue(s), in addition to the similarity of ol-2 and mlo resistances for both genetic and phytopathological characteristics, suggests that ol-2 is likely a mlo-homologue. Thus, a homology-based cloning approach could be more suitable than positional cloning for ol-2 isolation.     

籼稻多蘖矮半矮秆基因的遗传分析和基因定位

对籼稻标记基因系材料多蘖矮的遗传分析表明, 其矮生性状是由2对隐性半矮秆基因控制的,分别为sd1和一个新的半矮秆基因,该基因初步定名为sdt3。以多蘖矮与南京6号杂交F₂的分离群体为基础,应用SSR标记进行连锁分析,将半矮秆基因sdt3定位于第11染色体的SSR标记SSR98和SSR35之间,分别相距0.06 cM、0.13 cM,二者之间的物理距离约为93kb。以南京6号为轮回亲本与多蘖矮进行回交和自交获得由半矮秆基因sdt3控制的近等基因系（新多蘖矮）,以赤霉素处理表明由sdt3控制的半矮秆系新多蘖矮对赤霉素不敏感。