水稻纹枯病抗性关联分析及抗性等位变异发掘

采用苗期微室接种鉴定法,用144个分布于水稻全基因组的多态性标记,利用TASSEL软件GLM (Q)、MLM (Q+K)和MLM (PCA+K) 3种模型对456份水稻材料组成的自然群体进行纹枯病抗性关联分析。结果发现,有13个标记位点至少在两种模型中均被检测到与纹枯病抗性显著关联,单个位点可解释表型变异的1.84%~8.42%;其中10个标记位点位于以往报道的连锁定位的抗纹枯病QTL附近,3个标记位点(RM1036、RM5371和RM7585)是未曾报道的新的抗病相关位点。抗性等位变异RM7585-150对纹枯病发病病级减效效应最大;有259份材料携带抗性等位变异RM5371-129,占供试材料总数的56.8%,只有26份材料携带抗性等位变异RM1036-82,占供试材料总数的5.7%。水稻纹枯病发病病级与其含有的抗性等位变异数量呈极显著的负相关。本研究结果将为水稻抗纹枯病分子标记辅助育种提供理论依据。     

Identification of novel quantitative trait loci associated with brown planthopper resistance in the rice landrace Salkathi

The brown planthopper (BPH) is a potent pest of rice in Asia and Southeast Asia. Host resistance has been found to be the most suitable alternative to manage the insect. But varietal resistance has been found to be short-lived. There has been a constant search for alternate resistance genes. We developed an F₈ recombinant inbred population for the BPH resistance gene in Salkathi, an indica landrace from Odisha, India. Phenotyping of RILs against the BPH population at Cuttack, Odisha showed continuous skewed variation with four peaks at 2.1–3.0, 4.1–5.0, 6.1–7.0 and 8.1–9.0 SES score, suggesting the involvement of quantitative loci for resistance to BPH in Salkathi. Mapping showed the presence of two QTLs on the short arm of chromosome 4. One QTL, with phenotype variance of 37.02% is located between the markers RM551 and RM335. The other QTL, with phenotype variance of 7.1% is located between markers RM335 and RM5633. The two QTLs have been designated as qBph4.3 and qBph4.4. QBph4.3 seems to be a novel QTL associated with BPH resistance. We have successfully transferred qBph4.3 and qBph4.4 into two elite rice cultivars, Pusa 44 and Samba Mahsuri. Fine mapping of the identified QTLs may lead to a successful transfer of QTLs into other elite germplasm backgrounds.     

水稻抗白叶枯病新基因Xa32（t）的鉴定和初步定位

通过多菌系接种鉴定及抗谱分析,并与目前国际上已知抗白叶枯病基因比较,证明在水稻抗源C4064中含有一个新的抗白叶枯病基因,暂命名为Xa32(t)。应用分离集团分析法(BSA),借助SSR和EST等分子标记,对该基因进行了分子标记定位。通过对F₂分离群体及F₃家系单株进行遗传连锁性检测,发现6个位于水稻第11染色体长臂末端的分子标记RM27256、RM27274、RM2064、ZCK24、RM6293和RM5926与Xa32(t)基因连锁。它们与Xa32(t)基因间的遗传距离分别为2.1、1.0、1.0、0.5、1.5和2.6 cM。其中标记RM6293和RM5926位于染色体近端粒一侧,其他4个标记RM27256、RM27274、RM2064和ZCK24位于基因的另一侧。将Xa32(t)定位在水稻第11染色体长臂末端2.0 cM范围内。     

水稻抗白叶枯病新基因Xa32（t）的鉴定和初步定位

通过多菌系接种鉴定及抗谱分析,并与目前国际上已知抗白叶枯病基因比较,证明在水稻抗源C4064中含有一个新的抗白叶枯病基因,暂命名为Xa32(t)。应用分离集团分析法(BSA),借助SSR和EST等分子标记,对该基因进行了分子标记定位。通过对F₂分离群体及F₃家系单株进行遗传连锁性检测,发现6个位于水稻第11染色体长臂末端的分子标记RM27256、RM27274、RM2064、ZCK24、RM6293和RM5926与Xa32(t)基因连锁。它们与Xa32(t)基因间的遗传距离分别为2.1、1.0、1.0、0.5、1.5和2.6 cM。其中标记RM6293和RM5926位于染色体近端粒一侧,其他4个标记RM27256、RM27274、RM2064和ZCK24位于基因的另一侧。将Xa32(t)定位在水稻第11染色体长臂末端2.0 cM范围内。     

Resistance to rice sheath blight (<Emphasis Type="Italic">Rhizoctonia solani</Emphasis> Kühn) [(teleomorph: <Emphasis Type="Italic">Thanatephorus cucumeris</Emphasis> (A.B. Frank) Donk.] disease: current status and perspectives

Sheath blight (ShB) disease, caused by Rhizoctonia solani, is an economically important rice disease worldwide, especially in intensive production systems. Several studies have been conducted to identify sources for ShB resistance in different species of rice, including local accessions and landraces. To date, none of the genotypes screened are immune to ShB, although variation in levels of resistance have been reported. Several quantitative trait loci (QTL) for ShB resistance have been identified using mapping populations derived from indica or japonica rice. A total of 33 QTL associated with ShB resistance located on all 12 rice chromosomes have been reported, with ten of these co-localizing with QTL for morphological attributes, especially plant height, or for heading date. Sixteen QTL, from the same or differing genetic backgrounds, have been mapped at least twice. Of these, nine QTL were independent of morphological traits and heading date. We hypothesize that two main, distinct, mechanisms contribute to ShB resistance: physiological resistance and disease escape. Strategies to improve our understanding of the genetics of resistance to ShB are discussed.     

Identification of a major blast resistance gene in the rice cultivar 'Tetep'

Analysis of near‐isogenic lines (NILs) indicated the presence of a novel resistance gene in the indica rice cultivar ‘Tetep’ which was highly resistant to the rice blast fungus Magnaporthe grisea.‘Tetep’ was crossed to the widely used susceptible cultivar ‘CO39’ to generate the mapping population. A Mendelian segregation ratio of 3 : 1 for resistant to susceptible F₂ plants further confirmed the presence of a major dominant locus, in ‘Tetep’, conferring resistance to the blast fungal isolate B157, corresponding to the international race IC9. Simple sequence length polymorphism (SSLP) was used for molecular genetic analysis. The analysis revealed that the SSLP marker RM 246 was linked to a novel blast resistance gene designated Pi‐tp(t) in ‘Tetep’.     

Unveilingsources of stripe rust resistance in diverse wheat (<Emphasis Type="Italic">Triticum Aestivum</Emphasis> L.) germplasm using narrow down methodology: a proof of concept

Stripe rust of wheat caused by the fungal pathogen is a destructive foliar disease of wheat. Thus, it is crucial step to characterize the resistant germplasm for stripe rust in a diverse germplasm pool for their ultimate utilization in efficient crop rust resistance breeding. In the present study, we followed two pronged strategies involving integrated phenotypic and molecular characterization of 440 diverse wheat germplasm lines for rust resistance. The germplasm panel was extensively evaluated in field epiphytotic conditions during two consecutive years. After rigorous screening, 72 accessions were successfully revealed as resistant to moderately resistant to stripe rust. Subsequently, entries were then evaluated for their field agronomicperformances, considering prerequisites for serving as a donor germplasm,particularly for yield and 33 potential rust-resistant accessions were identified. Furthermore, to detect the sources of resistance, accessions were molecular characterized for potential race-specific resistance genes Yr5, Yr10,Yr15, and effective adult plant resistance (APR) gene Lr34/Yr18/pm38. We identified the 22 accessions possessing one or more single resistance genes and two accessions were observed with at least three of them. Moreover, Lr34/Yr18/pm38 was determined to confer resistance when observed along with any of the race-specific genes. Thus, the study not only provides proof of concept methodology to identify candidate resistant sources from large germplasm collections but simultaneouslyconfirmed the contribution of combining race-specific andnon-specific APR genes. The finding could further assist in the potential deployment of resistant genes directly into the stripe rust breeding program by involving marker-assisted approaches.     

Phenotypic and marker-assisted evaluation of spring and winter wheat germplasm for resistance to fusarium head blight

Fusarium head blight (FHB) caused by Fusarium species, is among the most devastating wheat diseases, causing losses in numerous sectors of the grain industry through yield and quality reduction, and the accumulation of poisonous mycotoxins. A germplasm collection of spring and winter wheat, including nine reference cultivars, was tested for Type II FHB resistance and deoxynivalenol (DON) content. Genetic diversity was evaluated on the basis of Simple Sequence Repeat (SSR) markers linked to FHB resistance quantitative trait loci (QTLs) and Diversity Arrays Technology (DArT) markers. The allele size of the SSR markers linked to FHB resistance QTLs from known resistance sources was compared to a germplasm collection to determine the presence of these QTLs and to identify potentially novel sources of resistance. Forty-two accessions were identified as resistant or moderately resistant to Fusarium spread, and two also had very low DON concentrations. Genetic relationships among wheat accessions were generally consistent with their geographic distribution and pedigree. SSR analysis revealed that several resistant accessions carried up to four of the tested QTLs. Resistant and moderately resistant lines without any known QTLs are considered to be novel sources of resistance that could be used for further genetic studies.     

QTL mapping of sheath blight resistance in a deep-water rice cultivar

Sheath blight, caused by the pathogen Rhizoctonia solani Kühn, is one of the most serious diseases of rice and leads to severe yield loss worldwide. A recombinant inbred line (RIL) population consisting of 121 lines was constructed from a cross between HH1B and RSB03, the latter of which is a deep-water rice variety. Five traits were used to evaluate sheath blight resistance, namely disease rating (DR), lesion length (LL), lesion height (LH), relative lesion length [RLL, the ratio of LL to plant height (PH)], and relative LH (RLH, the ratio of LH to PH). Using the RIL population and 123 molecular markers, we identified 28 quantitative trait loci (QTLs) for the five traits in two environments. These QTLs are located on nine chromosomes and most of them are environment specific. A major QTL for DR (qSBR1) on chromosome 1 was identified with contributions of 12.7% at Shanghai and 42.6% at Hainan, and it collocated with a QTL for PH. The allele at this locus from RSB03 enhances sheath blight resistance and increases PH. Another QTL for DR on chromosome 7 was adjacent to QTLs for heading date (HD) and four other disease traits. RSB03 also carries the resistant allele at this locus and shortens HD. The susceptible parent, HH1B, provides the resistance allele at the locus qSBR8, where QTLs for four other disease traits were identified. QTL mapping results showed that most QTLs for LL, LH, RLL, and RLH are collocated with QTLs for DR. Three QTLs for DR are independent from HD, PH, and four other disease traits, while four QTLs are closely related to HD and PH. Four QTLs for LL, LH, RLL, and RLH are independent from DR, HD, and PH, while there is only one region harboring QTLs for these four traits and HD. Correlation analysis and QTL mapping results indicated that LL, LH, RLL, and RLH might be important indices, like DR, for evaluating the level of resistance to rice sheath blight.     

Flanking SSR markers for allelism test for the Asian rice gall midge (Orseolia oryzae) resistance genes

Screening of rice germplasm against Asian rice gall midge, Orseolia oryzae (Wood-Mason), biotypes in India has led to identification of over 300 resistant rice genotypes. However, only ten resistance genes have been characterized so far. Identification of new genes through classical allelism test is tedious and time consuming. We propose to use closely linked flanking Simple Sequence Repeat (SSR) markers in allelism tests for identification of resistance genes. Of the ten known gall midge resistance genes, eight have been tagged and mapped. The Gm1 and Gm2 genes have closely linked flanking markers. Hence SSR markers RM219 and RM444, flanking the gene Gm1, and RM317, RM241 along with the SCAR marker F8, flanking the gene Gm2, were selected for this study. Tests with one set of 13 genotypes likely to carry Gm1 and another set of 17 genotypes suspected to contain Gm2 suggested the presence of the respective allele in all the 13 and 15 genotypes from these sets, respectively. Classical allelism test perfectly matched with the markers test. There were two exceptions involving amplification with RM444 in cultivar Kavya and with RM241 in genotype AE20, suggesting a single recombination which could have resulted in the mismatch. All the three markers in the genotype Bhumansan and the two flanking markers RM317 and F8 in AE20 indicated the absence of the Gm2 allele. This was validated through a classical test, revealing a segregation ratio of 15 resistant: 1 susceptible F₂ progeny of both the crosses between the Gm2 source Phalguna and these genotypes. We performed the allelism test with the markers on another set of 56 randomly selected gall midge resistant genotypes to discover possible sources of new resistance genes.     

Identification of SNP for rice blast resistance gene <Emphasis Type="Italic">Pike</Emphasis> and development of the gene-specific markers

Rice blast disease caused by Magnaporthe oryzae is an important limiting factor to rice production in the world. Introgression of blast resistance genes into improved germplasm by marker-assisted selection has been considered as an effective and environmentally beneficial means to control this disease. Pike, a broad-spectrum blast resistance gene, was cloned by map-based strategy recently in our laboratory. Two adjacent CC-NBS-LRR genes (designated as Pike-1 and Pike-2) were required for Pike-mediated resistance. In the current study, sequence alignment of the SNP G1328C and the SNP-surrounding region let us find that the Pik DNA variants of the studied rice lines appear to be divided into G-, C-, T- and G’-types. Based on the four genotypes, a Pike-specific marker system consisting of three PCR-based markers CP-G1328C, CP-G1328T and CP-G1328G’ was developed and used to effectively differentiate G-type allele from each of the others. Using this marker system, we investigated distribution of the Pik DNA variants in a set of 326 rice varieties or breeding lines and found that there were 2, 130, 135 and 59 rice lines identified to carry G-, C-, T- and G’-type alleles, respectively. In addition, with sequence data of the SNP G1328C-containing genomic region derived from 56 rice lines, we constructed a phylogenetic tree with three major clades which just corresponded to the types of the Pik DNA variants described above.     

Mining of favorable marker alleles for flag leaf inclination in some rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) accessions by association mapping

Flag leaf angle (FLA) in rice (Oryza sativa L.) is one of the important traits affecting F₁ seed production by mechanization. To elucidate the genetic mechanism of FLA and mine favorable marker alleles for F₁ seed production in rice, we performed a genome-wide association study using phenotypic data over 2 years and genotypic data of 262 pairs of simple sequence repeat (SSR) markers collected from 441 rice accessions. We detected seven SSR marker loci associated with FLA and four loci were novel. The four newly found loci were RM6266 on chromosome 3, RM348 on chromosome 4, RM258 on chromosome 10 and RM7303 on chromosome 11. We found a total of 27 favorable alleles, of which four, i.e., RM348-130 bp, RM7303-90 bp, RM258-180 bp, and RM4835-230 bp, had phenotypic effects larger than 10°. Nine combinations, which increased FLA by 45.7°–94.7° through pyramiding the favorable alleles contained in seven typical accessions, were predicted.     

Analysis of a major rice blast resistance gene in the rice restorer line Hanghui 1179

Rice blast, caused by the fungal pathogen Magnaporthe oryzae, is one of the most devastating diseases of rice (Oryza sativa) worldwide. Identification and utilization of resistance genes in rice breeding is considered to be an effective and economical method to control this disease. Hanghui 1179 (HH1179) is a new native rice restorer line developed in South China. The hybrids derived from HH1179 show broad-spectrum resistance against rice blast in South China, and a further understanding of the genetic resistance in HH1179 will provide useful information for breeding resistant cultivars. In the present study, we used bulked segregant analysis combined with specific-length amplified fragment sequencing to identify a dominant gene from HH1179 that provides resistance against the rice blast isolate GD13-14. Association analysis indicated that the resistance gene is located on chromosome 6 and we mapped the target gene to a 100.8 kb region (between markers InDel-8 and RM19818) that contains the Pi2/Pi9/Piz/Piz-t/Pi50 gene cluster. Candidate gene prediction and cDNA sequencing indicated that the target resistance gene in HH1179 is Pi2. Our findings will be valuable for resistance breeding with restorer line HH1179.     

Identification of a diverse mini‐core panel of Indian rice germplasm based on genotyping using microsatellite markers

Identification of a small core germplasm set representing the available genetic diversity is essential for its proper evaluation and subsequent utilization in rice improvement programmes. For constituting a small diverse mini‐core panel of Indian rice germplasm, a representative set of 6912 accessions drawn based on their geographic origin from the whole rice germplasm collection available in the National Gene Bank was genotyped using 36 microsatellite markers. Automated fragment analysis of amplicons yielded a total of 435 alleles, with an average 12.4 and range of 3–29 alleles per locus. Polymorphism information content (PIC) ranged from 0.08 (RGNMS190) to 0.86 (RM552) with an average of 0.528. Based on genotyping data, a mini‐core consisting of 98 genotypes was identified. Ninety‐four per cent of the alleles present in the core set were present in the mini‐core. The identified small but diverse panel will be useful for further intensive trait‐specific evaluation and utilization in allele mining.     

水稻抗纹枯病QTL表达的遗传背景及环境效应

利用水稻纹枯病菌强致病菌系RH-9人工接种Lemont导入到特青背景的213个近等基因导入系(TQ-ILs)群体和特青导入到Lemont背景的195个近等基因导入系(LT-ILs)群体,定位和分析了水稻抗纹枯病数量性状座位(quantitative trait loci, QTL)及其表达的环境与遗传背景效应。亲本Lemont对RH-9表现为高度感病,特青表现为中等抗病。人工接种后TQ-ILs群体的相对病斑高度(病斑高度与株高比)呈连续正态分布,LT-IL群体则明显偏向感病亲本Lemont。在不同年份和遗传背景下检测到影响纹枯病相对病斑高度的主效QTL 10个和互作QTL 13个,其中2006年在TQ-IL群体定位到的6个主效QTL在2007年均得到验证,表明这些QTL具有较好年度间的重复性。QSh4是唯一在双向导入系背景下表达的QTL,该位点特青等位基因降低相对病斑高度,提高抗性水平。在TQ-ILs群体中定位到位于第10染色体RM216~RM311区间的QSb10a与在LT-IL群体中定位到的位于相邻区间RM222~RM216的QSb10b的基因作用方向不同,推断这两个QTL存在紧密连锁关系。绝大多数在TQ-IL群体中表达的主效及互作QTL在LT-ILs群体中不表达,表明水稻抗纹枯病QTL具有明显的遗传背景效应。通过比较作图,本研究定位到的其中8个QTL在以往不同群体中同样被检测到,这些主效QTL对通过分子标记辅助选择(marker-assisted selection, MAS)培育水稻抗纹枯病育种可能具有应用价值。研究指出,标记辅助选择在不同遗传背景中能稳定表达的QTL或通过聚合不同抗病QTL是进一步提高水稻纹枯病抗性水平的一个有效途径。     

Detection of quantitative trait locus for leaffolder (Cnaphalocrocis medinalis (Guenée)) resistance in rice on linkage group 1 based on damage score and flag leaf width

Rice leaffolder (RLF) (Cnaphalocrocis medinalis (Guenée) is a destructive and widespread insect pest throughout the rice growing regions in Asia. The genetics of resistance to RLF in rice is very complex and not thoroughly explored. The present study was conducted to detect the quantitative trait loci (QTL) associated with RLF resistance involving 176 recombinant inbred lines (RILs) of F₈ generation derived from a cross between IR36, a leaffolder susceptible variety and TNAULFR831311, a moderately resistant indica rice culture. Simple sequence repeat (SSR) markers were used to construct specific linkage groups of rice. All the RILs were screened to assess their level of resistance to RLF by measuring the leaf area damaged. Besides this, the length and width of the flag leaf of each RIL were measured since these two parameters were considered as correlated traits to the RLF resistance in rice. All the above parameters observed across the RILs showed quantitative variation. Correlation analysis revealed that damage score based on greenhouse screening was positively correlated with length and width of the flag leaf. Out of 364 SSR markers analysed, 90 were polymorphic between the parents. Multi-point analysis carried out on segregating 69 SSR marker loci linkage group wise resulted in construction of linkage map with eleven groups of 42 SSR markers. Through single marker analysis, 19 SSR markers were found to have putative association with the three phenotypic traits studied. Of these markers, RM472 was identified as a locus having major effect on RLF resistance trait based on length of the flag leaf. Interval mapping detected two QTLs on linkage group 1. Among these QTLs, the QTL flanked by RM576–RM3412 were found to be associated with width of the flag leaf and RLF resistance. The putative SSR markers associated with leaffolder resistance identified in the present study may be one of the loci contributing resistance to RLF in rice.     

Further QTL mapping for yield component traits using introgression lines in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) under drought field environments

To better understand the underlying mechanisms of agronomic traits related to drought resistance and discover candidate genes or chromosome segments for drought-tolerant rice breeding, a fundamental introgression population, BC₃, derived from the backcross of local upland rice cv. Haogelao (donor parent) and super yield lowland rice cv. Shennong265 (recurrent parent) had been constructed before 2006. Previous quantitative trait locus (QTL) mapping results using 180 and 94 BC₃F_6,7 rice introgression lines (ILs) with 187 and 130 simple sequence repeat (SSR) markers for agronomy and physiology traits under drought in the field have been reported in 2009 and 2012, respectively. In this report, we conducted further QTL mapping for grain yield component traits under water-stressed (WS) and well-watered (WW) field conditions during 3 years (2012, 2013 and 2014). We used 62 SSR markers, 41 of which were newly screened, and 492 BC₄F_2,4 core lines derived from the fourth backcross between D123, an elite drought-tolerant IL (BC₃F₇), and Shennong265. Under WS conditions, a total of 19 QTLs were detected, all of which were associated with the new SSRs. Each QTL was only identified in 1 year and one site except for qPL-12-1 and qPL-5, which additively increased panicle length under drought stress. qPL-12-1 was detected in 2013 between new marker RM1337 and old marker RM3455 (34.39 cM) and was a major QTL with high reliability and 15.36% phenotypic variance. qPL-5 was a minor QTL detected in 2013 and 2014 between new marker RM5693 and old marker RM3476. Two QTLs for plant height (qPHL-3-1 and qPHP-12) were detected under both WS and WW conditions in 1 year and one site. qPHL-3-1, a major QTL from Shennong265 for decreasing plant height of leaf located on chromosome 3 between two new markers, explained 22.57% of phenotypic variation with high reliability under WS conditions. On the contrary, qPHP-12 was a minor QTL for increasing plant height of panicle from Haogelao on chromosome 12. Except for these two QTLs, all other 17 QTLs mapped under WS conditions were not mapped under WW conditions; thus, they were all related to drought tolerance. Thirteen QTLs mapped from Haogelao under WS conditions showed improved drought tolerance. However, a major QTL for delayed heading date from Shennong265, qDHD-12, enhanced drought tolerance, was located on chromosome 12 between new marker RM1337 and old marker RM3455 (11.11 cM), explained 21.84% of phenotypic variance and showed a negative additive effect (shortening delay days under WS compared with WW). Importantly, chromosome 12 was enriched with seven QTLs, five of which, including major qDHD-12, congregated near new marker RM1337. In addition, four of the seven QTLs improved drought resistance and were located between RM1337 and RM3455, including three minor QTLs from Haogelao for thousand kernel weight, tiller number and panicle length, respectively, and the major QTL qDHD-12 from Shennong265. These results strongly suggested that the newly screened RM1337 marker may be used for marker-assisted selection (MAS) in drought-tolerant rice breeding and that there is a pleiotropic gene or cluster of genes linked to drought tolerance. Another major QTL (qTKW-1-2) for increasing thousand kernel weight from Haogelao was also identified under WW conditions. These results are helpful for MAS in rice breeding and drought-resistant gene cloning.     

Identification of simple sequence repeat markers for sweetpotato weevil resistance

The development of sweetpotato [Ipomoea batatas (L.) Lam] germplasm with resistance to sweetpotato weevil (SPW) requires an understanding of the biochemical and genetic mechanisms of resistance to optimize crop resistance. The African sweetpotato landrace, ‘New Kawogo’, was reported to be moderately resistant to two species of SPW, Cylas puncticollis and Cylas brunneus. Resistance has been associated with the presence of hydroxycinnamic acids esters (HCAs), but the underlying genetic basis remains unknown. To determine the genetic basis of this resistance, a bi-parental sweetpotato population from a cross between the moderately resistant, white-fleshed ‘New Kawogo’ and the highly susceptible, orange-fleshed North American variety ‘Beauregard’ was evaluated for SPW resistance and genotyped with simple sequence repeat (SSR) markers to identify weevil resistance loci. SPW resistance was measured on the basis of field storage root SPW damage severity and total HCA ester concentrations. Moderate broad sense heritability (H² = 0.49) was observed for weevil resistance in the population. Mean genotype SPW severity scores ranged from 1.0 to 9.0 and 25 progeny exhibited transgressive segregation for SPW resistance. Mean genotype total HCA ester concentrations were significantly different (P < 0.0001). A weak but significant correlation (r = 0.103, P = 0.015) was observed between total HCA ester concentration and SPW severity. A total of five and seven SSR markers were associated with field SPW severity and total HCA ester concentration, respectively. Markers IBS11, IbE5 and IbJ544b showed significant association with both field and HCA-based resistance, representing potential markers for the development of SPW resistant sweetpotato cultivars.     

A new rice gall midge resistance gene in the breeding line CR57-MR1523, mapping with flanking markers and development of NILs

Gall midge is the third most destructive insect pests of rice after stem borers and planthoppers. Host plant resistance has been recognized as the most effective and economic, means for gall midge management. With the characterization of a new gall midge biotype (GMB) 4M, unique feature of gall midge resistance in the breeding line CR57-MR1523 was highlighted. Multi-location evaluation of F₃ families derived from the cross TN1 × CR57-MR1523 against different gall midge biotypes helped to identify a new dominant gene conferring resistance against GMB4. This gene has been designated as Gm11t. Though CR57-MR1523 has been extensively used in breeding gall midge resistant rice varieties like Suraksha, neither the genetics of resistance nor chromosomal location of the resistance gene(s) is known. In the present study we have tagged and mapped the new gall midge resistance gene, Gm11t, on chromosome 12, using SSR markers. To map the gene locus, 466 F₁₀ generation Recurrent Inbred Lines (RILs), from the cross of TN1 × CR57-MR1523 were used. Of the 471 SSR markers spread across the rice genome, 56 markers showed polymorphism and were used to screen a subset of the mapping population consisting of 10 resistant (R) and 10 susceptible (S) F₁₀ RILs. Six SSR markers, RM28706, RM235, RM17, RM28784, RM28574 and RM28564 on chromosome 12 were initially found to be associated with resistance and susceptibility. Based on the linkage analysis in selected 158 RILs, we were able to map the locus between two flanking SSR markers, RM28574 and RM28706, on chromosome 12 within 4.4 and 3.8 cM, respectively. Further, two NILs with 99% genetic similarity, were identified from the RILs which differed in gall midge resistance. The tightly linked flanking SSR markers will facilitate marker-assisted gene pyramiding and map-based cloning of the resistant gene. NILs would be valuable materials for functional analysis of the identified candidate gene.     

A relative expression of Xa7 gene controlling bacterial leaf blight resistance in indonesian local rice population (Oryza sativa L.)

Discovery of new alleles at important gene loci through allele mining could support the rice improvement program to sustain national food security. Evaluation of the existing Indonesian local rice landraces is an important point to detect the potential of functional alleles. One of the bacterial leaf blight (BLB) resistance alleles, Xa7 was detected in Indonesian rice landrace germplasm, Parekaligolara. To validate this potential allele, field evaluation on the segregating population, expression analysis using real time RT-PCR, and sequencing were carried out. Two selected Parekaligolara progenies lines (F₄ and F₆) from double crosses with other selected landraces were clearly more resistant to a dominant Indonesian BLB, Race IV. Specific primers of Xa7-LD40 successfully amplified the alleles of F₄ and F₆ lines approximately 300 bp in length. The amplicon sequenced using vector-targeted primers, resulting 264 bp which were flanked between 602 and 866 bp sites. The translated sequence which produced 60 amino acids (open reading frame) ORF, showed homology with the encoding gene associated with the defense system to biotic stress, BTB/POZ. As integrated researches for many potential biotic and abiotic stresses alleles on Indonesian landraces germplasm, this outcome expectedly supports rice landraces utilization for developing of elite cultivars which survive on global changed conditions and benefiting to national food security.