<Emphasis Type="Italic">Agrobacterium</Emphasis> mediated transformation of indica rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) for insect resistance

The rice leaffolder (RLF), Cnaphalocrocis medinalis is an important pest of rice that causes severe damage in many areas of the world. The plants were transformed with fully modified (plant codon optimized) synthetic Cry1C coding sequences as well as with the hpt and gus genes, coding for hygromycin phosphotransferase and β-glucuronidase, respectively. Cry1C sequences placed under the control of doubled 35S promoter plus the AMV leader sequence, and hpt and gus genes driven by cauliflower mosaic virus 35S promoter, were used in this study. Embryogenic calli after cocultivation with Agrobacterium were selected on the medium containing hygromycin B. A total of 67 hygromycin-resistant plants were regenerated. PCR and Southern blot analyses of primary transformants revealed the stable integration of Cry1C coding sequences into the rice genome with predominant single copy integration. R₁ progeny plants disclosed a monogenic pattern (3:1) of transgene segregation as confirmed by molecular analyses. These transgenic lines were highly resistant to rice leaffolder (RLF), Cnaphalocrocis medinalis as revealed by insect bioassay.     

Fine mapping of a <Emphasis Type="Italic">Xantha</Emphasis> mutation in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The recessive mutation of the XANTHA gene (XNT) transforms seedlings and plants into a yellow color, visually distinguishable from normal (green) rice. Thus, it has been introduced into male sterile lines as a distinct marker for rapidly testing and efficiently increasing varietal purity in seed and paddy production of hybrid rice. To identify closely linked markers and eventually isolate the XNT gene, two mapping populations were developed by crossing the xantha mutant line Huangyu B (indica) with two wild type japonica varieties; a total of 1,720 mutant type F₂ individuals were analyzed for fine mapping using polymorphic InDel markers and high dense microsatellite markers. The XNT gene was mapped on chromosome 11, within in a fragment of ~100 kb, where 13 genes are annotated. The NP_001067671.1 gene within the delimited region is likely to be a candidate XNT gene, since it encodes ATP-dependent chloroplast protease ATP-binding subunit clp A. However, no sequence differences were observed between the mutant and its parent. Bioinformatics analysis demonstrated that four chlorophyll deficient mutations that were previously mapped on the same chromosome are located outside the XNT region, indicating XNT is a new gene. The results provide useful DNA markers not only for marker assisted selection of the xantha trait but also its eventual cloning.     

Identification and characteristics of quantitative trait locus for grain protein content, <Emphasis Type="Italic">TGP12</Emphasis>, in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Grain protein content is an important analysis target to determine grain quality in rice. This study analyzed quantitative trait loci (QTLs) for the content of grain protein and amylose using the chromosomal segment substitution lines developed from ‘Koshihikari’ and ‘Nona Bokra’. It also evaluated the effects of target QTL on eating and cooking quality through the physical properties of cooked rice and its gel consistency. QTL analysis over 3 years detected the QTL on chromosome 12, TGP12, which consistently decreased total grain protein content via the ‘Nona Bokra’ allele. Selected CSSL with TGP12, CSSL-TGP12, showed a lower content of total grain protein in brown and milled rice, and had similar amylose content, grain size, and weight of brown rice, compared with ‘Koshihikari’. Based on the results of sodium dodecyl sulfate polyacrylamide gel electrophoresis, brown rice with CSSL-TGP12 had no remarkable decrease or loss in any specific protein. Regarding eating and cooking quality, CSSL-TGP12 did not show stable effects on physical properties, hardness, stickiness, or adherability of cooked rice or its gel consistency. These results suggest that TGP12 could be one of the key genetic factors for the alteration of grain protein content without an effect on eating or cooking quality.     

Identification of quantitative trait loci associated with drought tolerance traits in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) under PEG and field drought stress

Two recombinant inbred line F₁₀ rice populations (IAPAR-9/Akihikari and IAPAR-9/Liaoyan241) were used to identify quantitative trait loci (QTLs) for ten drought tolerance traits at the budding and early seedling stage under polyethylene glycol-induced drought stress, and two traits of leaf rolling index (LRI) and leaf withering degree (LWD) under field drought stress. The results showed that the drought-tolerance capacity of IAPAR-9 was stronger than that of Akihikari and Liaoyan241. Thirty-four QTLs for 12 drought tolerance traits were detected, and among them, in the IAPAR-9/Akihikari population, qLRI9-1 and qLRI10-1 for LRI were repeatedly detected in RM3600-RM553 on chromosome 9 and in RM6100-RM3773 on chromosome 10, respectively, at two times points of July 31 and August 13 in 2014. The two QTLs are stable against the environmental impact, and qLRI9-1 and qLRI10-1 explained 6.77–13.66% and 5.01–8.32% of the phenotypic variance, respectively, at the two times points. qLWD9-2 for LWD in the IAPAR-9/Liaoyan241 population contributed 8.73% of variation was detected in the same marker interval with the qLRI9-1, and qLRI1-1 for LRI and qLWD1-1 for LWD were located in the same marker interval RM11054-RM5646 on chromosome 1, which contributed 18.82 and 5.78% of phenotype variation respectively. qGV3 for germination vigor and qRGV3 for relative germination vigor at the budding stage were detected in the same marker interval RM426-RM570 on chromosome 3, which explained 14.98 and 16.30% of the observed phenotypic variation respectively, representing major QTLs. The above-mentioned stable or major QTLs regions could be useful for molecular marker assisted selection breeding, fine mapping, and cloning.     

A novel blast resistance locus in a rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) cultivar,Chumroo, of Bhutan

The rice cultivar ‘Chumroo’ is commonly cultivated in the mid- and high-altitude areas of Bhutan. This cultivar has shown durable blast resistance in that area, without evidence of breakdown, for over 20 years. Chumroo was inoculated with 22 blast isolates selected from the race differential standard set of Japan. The cultivar showed resistance to all the isolates. To identify the resistance gene(s), Chumroo was crossed with a susceptible rice cultivar, Koshihikari. The F₁ plants of the cross showed resistance. Segregation analyses of 300 F₃ family lines fitted the segregation ratio of 1:2:1 and indicated that a single dominant gene controls the resistance to a blast isolate Ao 92-06-2 (race 337.1). The Chumroo resistance locus (termed Pi46(t)) was mapped between two SSR markers, RM6748 and RM5473, on the terminal region of the long arm of chromosome 4, using linkage analysis with SSR markers. The nearest marker, RM5473, was linked to the putative resistance locus at a map distance of 3.2 cM. At the chromosomal region, no true resistance genes were identified, whereas two field resistance genes were present. Therefore, we designated Pi46(t) as a novel blast resistance locus.     

Identification of quantitative trait loci involved in resistance to <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">syringae</Emphasis> in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.)

Pseudomonas syringae is the main pathogen responsible for bacterial blight disease in pea and can cause yield losses of 70%. P. syringae pv. pisi is prevalent in most countries but the importance of P. syringae pv. syringae (Psy) is increasing. Several sources of resistance to Psy have been identified but genetics of the resistance is unknown. In this study the inheritance of resistance to Psy was studied in the pea recombinant inbred line population P665 × ‘Messire’. Results suggest a polygenic control of the resistance and two quantitative trait loci (QTL) associated with resistance, Psy1 and Psy2, were identified. The QTL explained individually 22.2 and 8.6% of the phenotypic variation, respectively. In addition 21 SSR markers were included in the P665 × ‘Messire’ map, of which six had not been mapped on the pea genome in previous studies.     

Fine mapping of a major quantitative trait locus, <Emphasis Type="Italic">qFLL6.2</Emphasis>, controlling flag leaf length and yield traits in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Fine mapping of a quantitative trait locus, qFLL6.2, controlling flag leaf length (FLL) and yield traits in rice was conducted using four sets of near isogenic lines (NILs) that were developed from a common residual heterozygote at F₇ generation of the indica rice cross Zhenshan 97/Milyang 46. Each of the NIL sets consisted of 40 lines that are S₁ progenies of ten maternal homozygotes, ten paternal homozygotes, and 20 heterozygotes differing in a portion of the 1.19-Mb interval RM3414–RM6917 on the short arm of rice chromosome 6. Analysis of phenotypic differences among the three genotypic groups in each NIL set delimited qFLL6.2 to a 62.1-kb region flanked by simple sequence repeat marker RM3414 and sequence-tagged site marker Si2944. This QTL explained 52.73% of the phenotypic variance, and the Zhenshan 97 allele increased FLL by 2.40 cm. Based on data collected from homozygous lines of three of the NIL sets, qFLL6.2 was shown to have major effects on all the three yield traits analyzed, including the number of spikelets per panicle, the number of filled grains per panicle, and grain weight per panicle. A comparison of the different groups revealed that the effect of qFLL6.2 was highly consistent across different genetic backgrounds and environments, providing a good candidate for map-based cloning and investigating the source–sink relationship in rice.     

RDA derived <Emphasis Type="Italic">Oryza minuta</Emphasis>-specific clones to probe genomic conservation across <Emphasis Type="Italic">Oryza</Emphasis> and introgression into rice (<Emphasis Type="Italic">O. sativa</Emphasis> L.)

Molecular markers have been successfully used in rice breeding however available markers based on Oryza sativa sequences are not efficient to monitor alien introgression from distant genomes of Oryza. We developed O. minuta (2n = 48, BBCC)-specific clones comprising of 105 clones (266–715 bp) from the initial library composed of 1,920 clones against O. sativa by representational difference analysis (RDA), a subtractive cloning method and validated through Southern blot hybridization. Chromosomal location of O. minuta-specific clones was identified by hybridization with the genomic DNA of eight monosomic alien additional lines (MAALs). The 37 clones were located either on chromosomes 6, 7, or 12. Different hybridization patterns between O. minuta-specific clones and wild species such as O. punctata, O. officinalis, O. rhizomatis, O. australiensis, and O. ridleyi were observed indicating conservation of the O. minuta fragments across Oryza spp. A highly repetitive clone, OmSC45 hybridized with O. minuta and O. australiensis (EE), and was found in 6,500 and 9,000 copies, respectively, suggesting an independent and exponential amplification of the fragment in both species during the evolution of Oryza. Hybridization of 105 O. minuta specific clones with BB- and CC-genome wild Oryza species resulted in the identification of 4 BB-genome-specific and 14 CC-genome-specific clones. OmSC45 was identified as a fragment of RIRE1, an LTR-retrotransposon. Furthermore this clone was introgressed from O. minuta into the advanced breeding lines of O. sativa.     

A simple,rapid and reliable bioassay for evaluating seedling vigor under submergence in <Emphasis Type="Italic">indica</Emphasis> and <Emphasis Type="Italic">japonica</Emphasis> rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Submergence is a major stress causing yield losses particularly in the direct-seeded rice cultivation system and necessitates the development of a simple, rapid and reliable bioassay for a large scale screening of rice germplasms with tolerance against submergence stress. We developed two new bioassay methods that were based primarily on the seedling vigor evaluated by the ability of fast shoot elongation under submerged conditions, and compared their effectiveness with two other available methods. All four bioassay methods using cultivars of 7 indica and 6 japonica types revealed significant and consistent cultivar differences in seedling vigor under submergence and/or submergence tolerance. Japonica cultivars were more vigorous than indica cultivars, with Nipponbare being the most vigorous. The simplest test tube method showed the highest correlations to all other methods. Our results suggest that seedling vigor serves as a submergence avoidance mechanism and confers tolerance on rice seedlings to flooding during early crop establishment. A possible relationship is discussed between seedling vigor based on fast shoot elongation and submergence tolerance defined by recovery from submergence stress.     

Mapping of quantitative trait loci controlling powdery mildew resistance in Mungbean (<Emphasis Type="Italic">Vigna radiata</Emphasis> (L.) Wilczek)

Powdery mildew disease in mungbean is caused by the fungus, Erysiphe polygoni D.C. We identified two quantitative trait loci (QTLs) controlling resistance to the disease in a RIL population of 190 F₇ lines. The population was developed from the cross between a susceptible cultivar, “Kamphaeng Saen 1” and a resistant line, “VC6468-11-1A”. Reaction to the disease was evaluated for resistance in field and greenhouse conditions. Results from analysis of variance revealed that 15 SSR loci on three linkage groups (LG) associated with the resistance. Composite interval mapping consistently identified two QTLs on two LGs, qPMR-1 and qPMR-2, conferring the resistance. qPMR-1 and qPMR-2 accounted for 20.10 and 57.81% of the total variation for plant response to the disease, respectively. Comparison based on common markers used in our and previous studies suggested that qPMR-2 is possibly the same as the major QTL reported earlier using another resistant source. The SSR markers flanking and closely linked to qPMR-1 (CEDG282 and CEDG191) and qPMR-2 (MB-SSR238 and CEDG166) are useful for marker-assisted selection for mungbean resistance to powdery mildew.     

Quantitative trait loci associated with seed set under high temperature stress at the flowering stage in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

High temperature stress (HTS), an increasingly important problem in rice production, significantly reduces rice yield by reducing seed set percentage (SSP). Breeding rice varieties with tolerance to HTS at the flowering stage is therefore essential for maintaining rice production as the climate continues to warm. In this study, two quantitative trait loci (QTL) underlying tolerance to HTS were identified using the recombinant inbred lines (RILs) derived from a cross between the HTS-tolerant rice cultivar 996 and the sensitive cultivar 4628. SSP was used as the heat-tolerance indicator for the lines, which were subjected to HTS at the flowering stage in both field and growth chamber experiments. Two major QTL that affected SSP in both conditions were detected in the interval between RM5687 and RM471 on chromosome 4, and between RM6132 and RM6100 on chromosome 10. The QTL located on chromosome 4 explained 21.3% in field and 25.8% in growth chamber of the total phenotypic variation in SSP, and increased the SSP of plants subjected to HTS by 9.1% in field and by 9.3% in growth chamber. The second QTL located on chromosome 10 explained 11.5% in field and 11.6% in growth chamber of the total phenotypic variation in SSP, and increased the SSP of plants subjected to HTS by 7.2% in field and 7.0% in growth chamber. The positive additive effects of the two QTL were derived from the 996 alleles. The two major QTL identified in this study could be useful for further fine mapping and cloning of these genes and for molecular marker-aided breeding of heat-tolerant rice cultivars.     

QTL Identification and Fine Mapping for Seed Storability in Rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Rice seed storability is an important characteristic of seed quality so that the cultivars with strong seed storability are expected in the production of hybrid seeds. Presently, little is known about the genetic and physiological mechanisms controlling rice seed storability. In this study, a double haploid population derived from the cross between a japonica cultivar CJ06 and an indica cultivar TN1 was used to identify the quantitative trait loci (QTLs) for seed germination percentage (GP) and fatty acid content (FA) during natural storage or artificial aging. A total of 19 QTLs, including ten QTLs for GP and nine QTLs for FA, were identified on nine chromosomes with the phenotypic variations ranged from 2.1 to 22.7%. Besides, six and three pairs of epistatic interactions were identified for GP and FA, respectively. Moreover, qGP-9, a QTL for germination percentage, was delimited in an interval of 92.8 kb between two STS markers P6 and P8, which contains 15 putative open reading frames. These results provide important information for understanding the genetic mechanisms on rice seed storability, and will be useful for breeding new rice varieties with high seed storability.     

Identification of quantitative trait loci for bruchid (<Emphasis Type="Italic">Callosobruchus maculatus</Emphasis>) resistance in black gram [<Emphasis Type="Italic">Vigna mungo</Emphasis> (L.) Hepper]

An inter-subspecific mapping population was generated by crossing V. mungo var. mungo (cv. TU 94-2, bruchid susceptible) and V. mungo var. silvestris (bruchid resistant). About 37.8% of the bruchid completed their lifecycle on seeds of V. mungo var. silvestris compared with 100% on the susceptible variety TU 94-2. The total developmental period of C. maculatus on Vigna mungo var. silvestris was considerably extended (88 days as compared with 34 days on TU 94-2). A genetic linkage map constructed using recombinant inbred lines (RILs) in F₉ generation with 428 markers [86 random amplified polymorphic DNA (RAPD), 47 simple sequence repeat (SSR), 41 inter-SSR (ISSR), 254 amplified fragment length polymorphism (AFLP)] was used for QTL detection. One hundred four individuals were used for detection of QTLs associated with bruchid resistance. The RILs exhibited a high level of variation in percentage adult emergence (0–100%) and developmental period (0–105 days). Two QTLs, Cmrae1.1 and Cmrae1.2, were identified for percentage adult emergence, on linkage group (LG) 3 and 4, respectively. For developmental period, six QTLs were identified, with two QTLs (Cmrdp1.1 and Cmrdp1.2) on LG 1, three QTLs (Cmrdp1.3, Cmrdp1.4, and Cmrdp1.5) on LG 2, and one QTL (Cmrdp1.6) on LG 10.     

Mapping of new resistance (<Emphasis Type="Italic">Vr2</Emphasis>, <Emphasis Type="Italic">Rm1</Emphasis>) and ornamental (<Emphasis Type="Italic">Di2</Emphasis>, <Emphasis Type="Italic">pl</Emphasis>) Mendelian trait loci in peach

Peach powdery mildew is one of the major diseases of the peach. Various sources of resistance to PPM have thus been identified, including the single dominant locus Vr2 carried by the peach rootstock ‘Pamirskij 5’. To map Vr2, a linkage map based on microsatellite markers was constructed from the F₂ progeny (WP²) derived from the cross ‘Weeping Flower Peach’ × ‘Pamirskij 5’. Self-pollinations of the parents were also performed. Under greenhouse conditions, all progenies were scored after artificial inoculations in two classes of reactions to PPM (resistant/susceptible). In addition to Vr2, WP² segregated for three other traits from ‘Weeping Flower Peach’: Rm1 for green peach aphid resistance, Di2 for double-flower and pl for weeping-growth habit. With their genomic locations unknown or underdocumented, all were phenotyped as Mendelian characters and mapped: Vr2 mapped at the top of LG8, at 3.3 cM, close to the CPSCT018 marker; Rm1 mapped at the bottom of LG1, at a position of 116.5 cM, cosegregating with the UDAp-467 marker and in the same region as Rm2 from ‘Rubira’^®; Di2 mapped at 28.8 cM on LG6, close to the MA027a marker; and pl mapped at 44.1 cM on LG3 between the MA039a and SSRLG3_16m46 markers. Furthermore, this study revealed, for the first time, a pseudo-linkage between two traits of the peach: Vr2 and the Gr locus, which controls the red/green color of foliage. The present work therefore constitutes a significant preliminary step for implementing marker-assisted selection for the four major traits targeted in this study.     

Mapping of QTLs associated with cadmium tolerance and accumulation during seedling stage in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Cadmium (Cd) is a non-essential element and toxic to plants. To investigate the genetics of Cd tolerance and accumulation in rice, quantitative trait loci (QTL) associated with Cd tolerance and accumulation at the seedling stage were mapped using a doubled haploid (DH) population derived from a cross between a japonica JX17 and an indica ZYQ8. A total of 22 QTLs were found to be associated with shoot height (SH), root length (RL), shoot dry weight (SDW), root dry weight (RDW), total dry weight (TDW) and chlorophyll content (CC), and 10 and 12 QTLs were identified under the control and Cd stress conditions, respectively. For Cd tolerant coefficient (CTC), 6 QTLs were detected on chromosomes 1, 3, 5, 8 and 10. Under Cd stress, 3 QTLs controlling root and shoot Cd concentrations were mapped on chromosome 6 and 7. One QTL for shoot/root rate of Cd concentration was identified on chromosome 3. The results indicated that Cd tolerance and accumulation were quantitatively inherited, and the detected QTLs may be useful for marker-assistant selection (MAS) and identification of the genes controlling Cd tolerance and accumulation in rice.     

Genetic control of agronomic traits in alfalfa (<Emphasis Type="Italic">M. sativa</Emphasis> ssp. <Emphasis Type="Italic">sativa</Emphasis> L.)

The objective of this study was to develop diallel population hybrids by crossing selected germplasm and to determine the gene effects and genetic control of yield and yield components using diallel analysis. A complete diallel including reciprocals was made during 2003 and 2004 between five alfalfa cultivars of different geographic origin. For each pairwise cross, five plants were chosen at random from each of the two cultivars (~100 florets per plant) to obtain the F₁ generation. A spaced plant field was established in 2006 which included the five alfalfa cultivars (parents) and their 20 diallel hybrids (F₁). The results of the diallel analysis suggest that the genetic control of major agronomic traits is determined by both additive gene action (accumulation of frequency of desirable alleles represented by significant GCA effects) and nonadditive gene action (complementary gene interactions represented by significant SCA effects). This type of gene action expression in alfalfa also determines the way in which breeding is carried out and brings about changes in the methods used and has given rise to the idea of the semi-hybrid breeding of this crop. The concept involves: breeding alfalfas within the population, identification of heterotic germplasm, and the production of seed of the population hybrid (PH).     

Sulfur alleviates cadmium toxicity in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) seedlings by altering antioxidant levels

We investigated changes in reactive oxygen species (ROS) and antioxidant levels in rice (Oryza sativa L. cv ‘Dongjin’) seedlings treated with toxic cadmium (Cd) and/or sulfur (S). Exposure of rice seedlings to 30 μM Cd inhibited plant growth and resulted in increased levels of superoxide, hydrogen peroxide, and malondialdehyde (MDA), and induced Cd uptake by the roots, stems, and leaves. Application of S to Cd-stressed seedlings ameliorated Cd-induced oxidative stress by increasing the capacity of the glutathione (GSH)-ascorbate (AsA) cycle, promoting S assimilation by increasing cysteine, GSH, and AsA content in treated plants, and decreasing Cd transfer from the roots to the stems and leaves. Therefore, these results indicate that S application represents a viable strategy of alleviating Cd-induced growth inhibition and oxidative damage by restricting Cd translocation from the roots to the stems and leaves, thereby maintaining sufficient levels of GSH and AsA by sustaining homeostasis of the GSH-AsA cycle.     

Mapping of quantitative trait loci controlling partial resistance against rust incited by <Emphasis Type="Italic">Uromyces pisi</Emphasis> (Pers.) Wint. in a <Emphasis Type="Italic">Pisum fulvum</Emphasis> L. intraspecific cross

A quantitative trait loci (QTL) associated with resistance to pea rust, caused by the fungus Uromyces pisi (Pers.) Wint., has been identified in a F₂ population derived from an intraspecific cross between two wild pea (Pisum fulvum L.) accessions, IFPI3260 (resistant) and IFPI3251 (susceptible). Both parental lines and all the segregating population displayed a fully compatible interaction (high infection type), which indicates absence of hypersensitive response. Nevertheless, differences on the percentage of symptomatic area of the whole plant (disease severity) were observed. A genetic map was developed covering 1283.3 cM and including 146 markers (144 random amplified polymorphic DNA (RAPDs) and two sequence tagged sites (STSs) markers) distributed in 9 linkage groups. A QTL explaining 63% of the total phenotypic variation was located in linkage group 3. RAPDs markers (OPY11₁₃₁₆ and OPV17₁₀₇₈) flanking this QTL should allow, after their conversion in SCARs, a reliable marker-assisted selection for rust resistance.     

Fine mapping of <Emphasis Type="Italic">Rf3</Emphasis> and <Emphasis Type="Italic">Rf4</Emphasis> fertility restorer loci of WA-CMS of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) and validation of the developed marker system for identification of restorer lines

The Wild Abortive (WA) system is the major cytoplasmic male sterility (CMS) source for hybrid rice production in indica rice and its fertility restoration is reported to be controlled by two major loci viz. Rf3 on chromosome 1 and Rf4 on chromosome 10. With the availability of the rice genome sequence, an attempt was made to fine map, develop candidate gene based markers for Rf3 and Rf4 and validate the developed marker system in a set of known restorer lines. Using polymorphic markers developed from microsatellite markers and candidate gene based markers from Rf3 and Rf4 loci, local linkage maps were constructed in two mapping populations of ~1,500 F₂ progeny from KRH2 (IR58025A/KMR3R) and DRRH2 (IR68897A/DR714-1-2R) hybrids. QTLs and their interactions for fertility restoration in Rf3 and Rf4 loci were identified. The identified QTL in both mapping populations together explained 66–72 % of the phenotypic variance of the trait suggesting their utility in developing a marker system for identification of fertility restorers for WA-CMS. Sequence comparison of the two candidate genes from the Rf3 and Rf4 regions in male sterile (A) and restorer (R) lines showed 2–3 bp indels and a few substitutions in the Rf3 region and indels of 327 and 106 bp in the Rf4 region respectively. The marker system identified in the present study was validated in 212 restorers and 34 maintainers along with earlier reported markers for fertility restoration of WA-CMS. Together DRCG-RF4-14 and DRCG-RF4-8 for the Rf4 locus and DRRM-RF3-5/DRRM-RF3-10 for the Rf3 locus showed a maximum efficiency of 92 % for identification of restorers.     

Participatory selection assisted by DNA markers for enhanced drought resistance and productivity in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

In this study, a F₂ population derived from the cross between deep-rooted variety “Moroberekan” with shallow-rooted variety “IR20” were used to identify and validate of SSR markers associated with root morphological traits. The F₂ lines were divided into two groups. In the first group, 152 seedling having minimum of four tillers were chosen and separated into four plantlets to plant them in polyvinyl chloride pipes for root study under well-watered (WW) condition at maturity stage. The lines were genotyped using SSR markers. QTLs for maximum root length (MRL) and root dry weight showed co-segregation with RM472, RM7 and RM201. The same material was forwarded to next generation (F₃) to validate the linked markers under both WW and low-moisture stress (LMS) conditions. These three markers were associated consistently with MRL across generations. In the second group, 1240 F₂ plants were forwarded to F₅ using SSD breeding method to test the effectiveness of the marker-assisted selection (MAS) method for drought resistant. The high performing genotypic group was significantly superior to low performing genotypic group for MRL, grain yield, root volume, root dry weight and root number, indicating the efficiency of MAS for root-related traits under LMS. Comparing MAS with farmer selection in F₆, the results showed that MAS group means were significantly different from farmer group means for MRL, root volume, root dry weight and root number. Thus, MAS was combined with participatory selection to select five high-yielding and deep rooted promising lines. Identification of stable QTL for root morphological traits under WW and LMS conditions can aid in MAS and to introduce them into varieties with good yield potential and accepted by farmer.