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.     

Genome-wide association analysis of seed germination percentage and germination index in <Emphasis Type="Italic">Brassica napus</Emphasis> L. under salt and drought stresses

Identifying genetic loci and possible candidate genes related to salt and drought stresses would be an economical, feasible and efficient way to accelerate the progress of abiotic tolerance breeding in rapeseed. In this study, a seed germination experiment using distilled water, salt and drought stresses was carried out with 520 B. napus germplasm resources. The rapeseed accessions showed large variation in the seed germination percentage (GP) and germination index (GI), with GP ranging from 26 to 100%, 0 to 100% and 0 to 100% and GI from 3.05 to 53.92, 0 to 24.50 and 0 to 25.00 in distilled water, under salt stress and drought stress, respectively. The coefficients of GP and GI were 0.72, 0.97 and 0.96 in distilled water, under salt stress and drought stress, respectively. A genome-wide association analysis of the 520 accessions was performed using the Q+K model and 31,839 single-nucleotide polymorphism (SNP) sites, revealing 23 SNPs associated with GP and 20 SNPs associated with GI under salt and drought stresses. The significant loci rs32406 and rs9466 on chromosome C08 and A04 were mapped to sites 3.32 and 2.20 kb from the salt- and drought-responsive gene BnaC08g41070 and drought-responsive gene BnaA04g02620D, respectively. BnaC06g01910D, a drought-responsive gene on chromosome C06, was found to overlap with the significant locus rs44427. In addition, 37 candidate genes associated with GP and GI under salt and drought stresses were detected. Most of these candidate genes regulate signal transduction and proline biosynthesis or encode a ubiquitin ligase E3; some have unknown biological functions and may respond to salt and drought stresses together as a complex network.     

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.     

Identification of drought responsive QTLs during vegetative growth stage of rice using a saturated GBS-based SNP linkage map

Drought is a major abiotic constraint for rice production worldwide. The quantitative trait loci (QTLs) for drought tolerance traits identified in earlier studies have large confidence intervals due to low density linkage maps. Further, these studies largely focused on the above ground traits. Therefore, this study aims to identify QTLs for root and shoot traits at the vegetative growth stage using a genotyping by sequencing (GBS) based saturated SNP linkage map. A recombinant inbred line (RIL) population from a cross between Cocodrie and N-22 was evaluated for eight morphological traits under drought stress. Drought was imposed to plants grown in 75 cm long plastic pots at the vegetative growth stage. Using a saturated SNP linkage map, 14 additive QTLs were identified for root length, shoot length, fresh root mass, fresh shoot mass, number of tillers, dry root mass, dry shoot mass, and root-shoot ratio. Majority of the drought responsive QTLs were located on chromosome 1. The expression of QTLs varied under stress and irrigated condition. Shoot length QTLs qSL1.38 and qSL1.11 were congruent to dry shoot mass QTL qDSM1.38 and dry root mass QTL qDRM1.11, respectively. Analysis of genes present within QTL confidence intervals revealed many potential candidate genes such as laccase, Calvin cycle protein, serine threonine protein kinase, heat shock protein, and WRKY protein. Another important gene, Brevis radix, present in the root length QTL region, was known to modulate root growth through cell proliferation and elongation. The candidate genes and the QTL information will be helpful for marker-assisted pyramiding to improve drought tolerance in rice.     

Overcoming unilateral incompatibility in crosses with wild allotetraploid potato species <Emphasis Type="Italic">Solanum stoloniferum</Emphasis> Schldtl. &amp; Bouchet

The germplasm of valuable for breeding wild allotetraploid potato species Solanum stoloniferum is rarely used because of pre- and postzygotic reproductive barriers with cultivated potatoes. One of the factors that complicate crosses between S. stoloniferum and S. tuberosum is unilateral incompatibility (UI). Here, we present the results of application of S. verrucosum and S _v S _v -lines for overcoming UI in crosses with S. stoloniferum and of generating male fertile hybrids derived from this species. S _v S _v -lines are F2 S. tuberosum dihaploid × S. verrucosum that are male fertile and have D/γ-type cytoplasm. Since they are homozygous for S _v gene from S. verrucosum, they were expected to have the same ability for elimination of prezygotic incompatibility as this species. Three accessions of S. verrucosum and seven S _v S _v -lines were pollinated by 26 accessions of S. stoloniferum. The crosses with S. verrucosum failed or had low efficacy (1.5–2.4 seeds per pollination). On the other hand, use of S _v S _v -lines was more efficient: 15.8 seeds per pollination. In spite of low percentage of germination (1.9%), 40 seedlings of interspecific hybrids were produced. The experiment on hybridization between S _v S _v -lines and S. stoloniferum has been reproduced with the accession PI 205522 of the wild species, which had DNA markers of PVY and LB resistance genes and W/γ cytoplasm: 950 hybrid seeds and 12 viable seedlings were produced. The genome of the seedlings was doubled by colchicine treatment, which generated hexaploids that formed highly fertile pollen and set seeds from self-pollination. We were able to cross them as females with the variety Katahdin.     

Identification and validation of root length QTLs for water stress resistance in hexaploid wheat (<Emphasis Type="Italic">Titicum aestivum</Emphasis> L.)

Thorough understanding of the genetic mechanisms governing drought adaptive traits can facilitate drought resistance improvement. This study was conducted to identify chromosome regions harbouring QTLs contributing for water stress resistance in wheat. A RIL mapping population derived from a cross between W7984 (Synthetic) and Opata 85 was phenotyped for root length and root dry weight under water stress and non-stress growing conditions. ANOVA showed highly significant (p ≤ 0.01) variation among the RILs for both traits. Root length and root dry weight showed positive and significant (p ≤ 0.01) phenotypic correlation. Broad sense heritability was 86% for root length under stress and 65% for root dry weight under non-stress conditions. A total of eight root length and five root dry weight QTLs were identified under both water conditions. Root length QTLs Qrln.uwa.1BL, Qrln.uwa.2DS, Qrln.uwa.5AL and Qrln.uwa.6AL combined explained 43% of phenotypic variation under non-stress condition. Opata was the source of favourable alleles for root length QTLs under non-stress condition except for Qrln.uwa.6AL. Four stress specific root length QTLs, Qrls.uwa.1AS, Qrls.uwa.3AL, Qrls.uwa.7BL.1 and Qrls.uwa.7BL.2 jointly explained 47% of phenotypic variation. Synthetic wheat contributed favourable alleles for Qrls.uwa.1AS and Qrls.uwa.3AL. Two stable root dry weight QTLs on chromosomes 4AL and 5AL were consistently found in both water conditions. Three validation populations were developed by crossing cultivars Lang, Yitpi, and Chara with Synthetic W7984 to transfer two of the QTLs identified under stress condition. The F_2.3 and F_3.4 validation lines were phenotyped under the same level of water stress as RILs to examine the effect of these QTLs. There were 13.5 and 14.5% increases in average root length due to the inheritance of Qrls.uwa.1AS and Qrls.uwa.3AL, respectively. The result indicated that closely linked SSR markers Xbarc148 (Qrls.uwa.1AS) and Xgwm391 (Qrls.uwa.3AL) can be incorporated into MAS for water stress improvement in wheat.     

Identification of QTLs associated with heat tolerance at the heading and flowering stage in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The ongoing rise in temperatures caused by global climate change is a critical climatic risk factor for rice production, and enhancing rice heat tolerance is an area of particular research interest. A recombinant inbred line (RIL) mapping population was developed from heat sensitive, rice cultivar IAPAR-9 crossed with heat tolerant, Liaoyan241. RIL and parental lines were exposed to high temperature at the heating and flowering stage in experiments in 2014 and 2015. As indicators of heat tolerance, the seed setting rate under natural (NS) and heat stress (HTS) conditions were measured, and the reduction rate of seed set (RRS) was calculated. Quantitative trait loci (QTL) analysis revealed eleven heat tolerance QTLs located on chromosomes 1, 3, 4, 5, and 6. Single QTL contribution rates were 4.75–13.81% and effect values were ? 5.98 to 5.00. Four major QTLs (qNS1, qNS4, qNS6, and qRRS1) were stable detected in different environments in both years. Thirteen QTLs with epistatic interactions and nine QTLs with environmental interactions were also detected. Major QTLs were all involved in epistatic and environmental interactions. Three QTLs from the SSR marker interval RM471 to RM177 region of chromosome 4 (qNS4, qHTS4, and qRRS4) were all involved in epistatic and environmental interactions and contributed to phenotypic variation, indicating that this region constituted a major QTL hotspot. The major QTL for heat tolerance identified in this study will aid in breeding tolerant cultivars and facilitating investigation of the molecular underpinnings of heat tolerance in rice.     

Identification of QTLs controlling low-temperature germinability and cold tolerance at the seedling stage in rice (<Emphasis Type="Italic">Oryza Sativa</Emphasis> L.)

Both low-temperature germinability (LTG) and cold tolerance at the seedling stage (CTS) are important traits for rice. In this study, a rice population of recombinant inbred lines (RILs), derived from the backcross population of a cross between Dongnong422 and Kongyu131, was developed to detect quantitative trait loci (QTL) affecting LTG and CTS by using seed of different storage times. Correlation analysis indicated that there was no significant relationship between LTG and CTS, suggesting that cold tolerance might be genetic differences for LTG and CTS. In total, Twelve and twenty-three major QTLs were detected for LTG and CTS, respectively, which could explain greater than 10% of the phenotypical variation. Eight (qCG12-1, qGI12-1, qGV9-1, qMLIT12-1, qPV6-1, qMDG12-1, qLDWcold10-1, qLFWcold10-1) significant QTLs were mapped for different storage time, it concluded that such QTLs were not affected by environment (storage time) and were closely related QTLs to cold tolerance. One or more QTLs were identified for each trait with some of these QTLs co-locating, qMLIT7-1, qCG7-1, and qGI7-1 for LTG, qLFWcold10-1, and qLDWcold10-1 for CTS with contributions over 15% were mapped common marker interval, respectively, co-location of QTLs for different traits can be an indication that a locus has pleiotropic effects on multiple traits due to a common mechanistic basis. Two lines, RIL128 and RIL73, might be valuable to improve the LTG and CTS through a combination of crosses. The identified QTLs might be applicable to improve the rice cold tolerance by the marker-assisted selection approach.     

Evaluation of morpho-physiological traits of MRQ74 pyramided lines with drought yield QTLs

Drought regularly affects rainfed lowland and upland rice ecosystems in Malaysia. Three drought yield QTLs, viz qDTY _2.2 , qDTY _3.1 and qDTY _12.1 successfully pyramided into MRQ74 to increase its yield under reproductive stage drought stress (RS). Forty-eight genotypes comprising 39 pyramided lines (PLs) with different qDTYs combinations, four parents including MRQ74 (recipient) and five checks were evaluated for morpho-physiological traits under RS and non-stress (NS). This study aims to determine which traits influenced by individual qDTY and qDTY combinations and to gain better understanding of QTL interactions in enhancing grain yield (GY) under RS. Results showed plant height, number of panicles, root length, root weight, relative water content and 100-grain weight increased while chlorophyll content and GY decreased under RS compared to NS. No significant difference was observed in days to flowering, leaf rolling and grain length between selected PLs and MRQ74 under RS. Six PLs with yield advantage (YA) of 208.17–1751.63 kg ha^?1 compared to MRQ74 in RS but yielded similar to MRQ74 under NS were further selected. Under RS, qDTY class analysis showed qDTY _12.1 individually and combination qDTY _12.1  + ?qDTY _2.2 produced the highest yield of 1521.77 and 1092.30 kg ha^?1 respectively. qDTY _12.1 as single or combination with other qDTY is the best qDTY in stabilizing GY under RS. PL-77 with qDTY _12.1 is the best PL with YA of more than 1100 kg ha^?1 compared to MRQ74 in both RS and NS conditions can be recommended for cultivation in normal and drought-prone areas.     

Morph-physiological responses of cotton interspecific chromosome substitution lines to low temperature and drought stresses

Limited knowledge about genetic and physiological traits associated with drought and low temperature stresses and narrow genetic diversity in Upland cotton (Gossypium hirsutum L.) are serious impediments in its genetic improvement. The objectives of this research were to determine the genetic and physiological traits associated with drought and low temperature effects and to identify chromosomal effects on these traits using chromosome substitution (CS) lines from three alien species of Gossypium, G. barbadense, G. tomentosum, and G. mustelinum, respectively. Two experiments were conducted to study low temperature and drought stress effects during seedling emergence and early growth stages in 21 cotton CS-lines with parent, Texas Marker (TM)-1. In Experiment I, plants were grown at optimum (30/22 °C) and low (22/14 °C) temperature conditions under optimum water and nutrient conditions. In Experiment II, plants were grown at optimum water (soil moisture content of 0.167 m³ m^?3) and in drought (soil moisture content 0.105 m³ m^?3) conditions under optimum temperature conditions. Above- and below-ground growth traits including several root traits of the CS lines were assessed at 25 days after sowing. The findings suggest which substituted chromosome or chromosome segment from the alien species likely harbors one or more genes for higher and lower tolerance to low temperature, respectively. CS-T04 and CSB08sh showed higher and lower tolerance to low temperature, respectively and CS-T04 and CS-B22sh showed higher and lower tolerance, respectively, to drought. CS lines are valuable analytical tool and useful genetic resources for targeted exploitation of beneficial genes for drought and low temperature stresses in Upland cotton.     

Development of salt tolerant IR64 near isogenic lines through marker-assisted breeding

Salt stress causes considerable damage to rice with a consequent reduction in grain yield, however, conventional breeding for this stress is time-consuming and costly. Recently, marker-assisted breeding has shown enormous potential to accelerate breeding of stress tolerant varieties because of its precision, time saving, and cost effectiveness. The present study was carried out to transfer Saltol, a major QTL on chromosome 1 associated with salinity tolerance, from FL478, a tolerant genotype, into IR64, a popular lowland variety through marker-assisted backcrossing (MABC). This technique considerably enhanced the recovery rate of the recurrent parent genome within three backcross generations, which could have saved several backcrosses compared with conventional schemes to achieve the same results. By using this technique, up to 99.7% of the recurrent parent genome was recovered at BC₃F₂ generation, saving at least three backcrosses compared with conventional breeding schemes. Salinity tolerance of IR64-Saltol lines was evaluated using saline culture solution adjusted to electrical conductivity of 12 dS m^-1 using NaCl. Based on selected physiological and growth parameters, the new Saltol introgression lines showed a significantly higher tolerance of salinity than their recurrent parent IR64. The results of this study confirm the benefits of using molecular markers in plant breeding to enhance tolerance of abiotic stresses.     

Genetic transformation of pigeonpea (<Emphasis Type="Italic">Cajanus cajan</Emphasis> L.) and screening transgenic progenies based on lateral root inhibition

Production of transgenic pigeonpea is becoming increasingly important, but the methods currently employed in production and subsequent screening still requires improvement. Here, we describe Agrobacterium-mediated genetic transformation of pigeonpea with reporter uidA (gus) gene and selectable marker, neomycin phospho-transferase (nptII) gene. Histochemical assay demonstrate localization of gus activity in cells and transformed plants. Overall, a transformation frequency of 0.33% was achieved using the protocol. Grafting of in vitro-regenerated healthy shoots indicates higher survival percent (72.6%), when stock and scion are of the same variety. Seeds harvested from primary transgenic plants can be screened based on lateral root inhibition strategy. Approximately 87% of the screened T₁ plants were found to be PCR positive. In conclusion, in vitro grafting of transgenic pigeonpea shoots leads to better plant establishment and screening based on lateral root inhibition leads to quick identification of positive segregants.     

Analysis of population structure and genetic diversity reveals gene flow and geographic patterns in cultivated rice (<Emphasis Type="Italic">O. sativa</Emphasis> and <Emphasis Type="Italic">O. glaberrima</Emphasis>) in West Africa

To fully exploit the diversity in African rice germplasm and to broaden the gene pool reliable information on the population genetic diversity and phenotypic characteristics is a prerequisite. In this paper, the population structure and genetic diversity of 42 cultivated African rice (Oryza spp.) accessions originating from West Africa (Benin, Mali and Nigeria, Liberia etc.) were investigated using 20 simple sequence repeats (SSR) and 77 amplified fragment length polymorphisms (AFLP). Additionally, field trials were set up to gain insight into phenotypic characteristics that differentiate the genetic populations among rice accessions. The analysis revealed considerably high polymorphisms for SSR markers (PIC mean?=?0.78) in the germplasm studied. A significant association was found between AFLP markers and geographic origin of rice accessions (R?=?0.72). Germplasm structure showed that Oryza sativa accessions were not totally isolated from Oryza glaberrima accessions. The results allowed identification of five O. glaberrima accessions which grouped together with O. sativa accessions, sharing common alleles of 18 loci out of the 20 SSR markers analyzed. Population structure analysis revealed existence of a gene flow between O. sativa and O. glaberrima rice accessions which can be used to combine several interesting traits in breeding programs. Further studies are needed to clarify the contributions of this gene flow to valuable traits such as abiotic and biotic stresses including disease resistance.     

Tissue culture and <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated genetic transformation studies in four commercially important indica rice cultivars

This research was undertaken to find an efficient tissue culture system and Agrobacterium-mediated genetic transformation method for recalcitrant indica rice cultivars. For this, mature seeds of commercially important indica rice varieties, ASD16, ADT43, IR 64, and Pusa Basmati were cultured on MS and N6 medium supplemented with 2 mg l^-1 2, 4-D + 30 g l^-1 sucrose. The calli grown in N6 medium showed better friability and embryogenic response. Out of the four varieties tested, ASD16 and IR64 showed better callusing and embryogenic capacity as compared to ADT43 and Pusa Basmati. For genetic transformation studies, embryogenic calli of all the cultivars were co-cultivated with the Agrobacterium tumefaciens strain LBA 4404 harboring the binary vector pCambia 1305.1 with GUS gene. GUS assay was performed for the putative transformed calli and its activity was found to be qualitatively higher in ASD16 and IR64 than the other two varieties. The best responsive ASD16 transformed calli was regenerated and the putative transgenic lines were regenerated. ASD16 transformed calli were confirmed by GUS assay. PCR analysis confirmed the presence of both GUS and HPT genes in ASD16 transgenic lines.     

Marker-assisted breeding to develop the drought-tolerant version of Sabitri,a popular variety from Nepal

Sabitri is a rice variety grown in a large part of the rainfed areas of Nepal. It was originally developed for irrigated condition; hence, this variety suffers high yield decline under drought. Two QTLs, qDTY _3.2 and qDTY _12.1 , with large effects on grain yield under drought were identified in the Sabitri background in separate QTL mapping studies. The present study reports the development of Sabitri near isogenic lines (NILs) with combinations of these two QTLs and their characterization under drought. To do so, marker-assisted backcross breeding (MABB) was combined with phenotypic selection to develop high-yielding drought-tolerant NILs with Sabitri grain type. Apart from this, drought-tolerant variants for grain type with high yield under non-stress were identified among the developed NILs. Early days to flowering of up to 13 days and reduction in plant height of up to 13 cm as compared to Sabitri were observed in the developed NILs. Some of these NILs showed higher yield compared to Sabitri and relatively higher tolerance to drought, indicating the capture of positive alleles and interactions during the course of selection. The developed NILs possessed high yield potential which make them suitable materials for the testing of water-saving technologies in irrigated areas. Based on their performance, these NILs can be deployed in rainfed areas in Nepal and other countries of South Asia to increase yield stability.     

Effects of introgressions from <Emphasis Type="Italic">Festuca pratensis</Emphasis> on winter hardiness of <Emphasis Type="Italic">Lolium perenne</Emphasis>

Previous studies reported that some genotypes with introgressed Festuca chromosome segment(s) in Lolium genome showed enhanced winter hardiness compared to Lolium. The aim of this study was to search comprehensively for the Festuca pratensis chromosome regions affecting winter hardiness-related traits when introgressed into the Lolium perenne genome. Association between F. pratensis introgression and winter hardiness-related traits (fall and winter hardiness indexes, early-spring dry matter yield, and freezing tolerance) were screened in the diploid introgression populations (n = 203) that had some F. pratensis chromosome segments introgressed. Eighty-four intron markers corresponding to unique rice genes randomly distributed across the genome were used for genotyping. Winter hardiness of almost all plants in the introgression populations was lower than that of the F. pratensis and triploid hybrid parents, but the average was higher than that of L. perenne. A significant positive effect of F. pratensis introgression on early-spring dry matter yield was detected on chromosome 7. This quantitative trait locus (QTL) was confirmed by linkage analysis using a backcross population with F. pratensis introgression in the target region of chromosome 7. However, the contribution of the newly identified QTL was rather small (6.7–9.6%), suggesting that superior winter hardiness of F. pratensis compared to L. perenne is conferred by multiple small-effect QTLs. We also detected a previously unreported negative effect of Festuca introgression on winter hardiness. Newly obtained QTL information in this study would contribute to the design of Festuca/Lolium hybrid breeding.     

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.     

Applications of polysaccharides (β-glucan) for physiological and biochemical parameters for evaluation rice tolerance under salinity stress at seedling stage

Salt stress is considered to be the most important abiotic stress and is the main problem in the interruption of the metabolic processes of plants, thereby limiting crop production. The present study was carried out to examine the effects of exogenously applied mushroom polysaccharides (β-glucan) on the growth of two rice varieties of Oryza sativa L. (MRQ74 and MR269) seedlings grown in vitro in 200 Mm sodium chloride (NaCl). Growth characteristics, including shoot and root height, and fresh and dry weight of salt-stressed rice seedlings were inhibited by sodium chloride treatment, the degree of which depends on the rice variety. The negative effects of salinity stress resulted in an imbalance of N, K⁺ Na⁺ and Na/K ratios and biochemical analysis activities a degradation of chlorophyll content in the plants. However, treatment with exogenously applied polysaccharides (β-glucan) ameliorated the stress generated by NaCl and improved the parameters mentioned above. The significance of the changes in growth and metabolism to salt stress tolerance in rice seedlings are discussed in the paper.     

Identification of genetic sources with attenuated <Emphasis Type="Italic">Tomato chlorosis virus</Emphasis>-induced symptoms in <Emphasis Type="Italic">Solanum</Emphasis> (section <Emphasis Type="Italic">Lycopersicon</Emphasis>) germplasm

The whitefly-transmitted Tomato chlorosis virus (ToCV) (genus Crinivirus) is associated with yield and quality losses in field and greenhouse-grown tomatoes (Solanum lycopersicum) in South America. Therefore, the search for sources of ToCV resistance/tolerance is a major breeding priority for this region. A germplasm of 33 Solanum (Lycopersicon) accessions (comprising cultivated and wild species) was evaluated for ToCV reaction in multi-year assays conducted under natural and experimental whitefly vector exposure in Uruguay and Brazil. Reaction to ToCV was assessed employing a symptom severity scale and systemic virus infection was evaluated via RT-PCR and/or molecular hybridization assays. A subgroup of accessions was also evaluated for whitefly reaction in two free-choice bioassays carried out in Uruguay (with Trialeurodes vaporariorum) and Brazil (with Bemisia tabaci Middle-East-Asia-Minor1—MEAM1?=?biotype B). The most stable sources of ToCV tolerance were identified in Solanum habrochaites PI 127827 (mild symptoms and low viral titers) and S. lycopersicum ‘LT05’ (mild symptoms but with high viral titers). These two accessions were efficiently colonized by both whitefly species, thus excluding the potential involvement of vector-resistance mechanisms. Other promising breeding sources were Solanum peruvianum (sensu lato) ‘CGO 6711’ (mild symptoms and low virus titers), Solanum chilense LA1967 (mild symptoms, but with high levels of B. tabaci MEAM1 oviposition) and Solanum pennellii LA0716 (intermediate symptoms and low level of B. tabaci MEAM1 oviposition). Additional studies are necessary to elucidate the genetic basis of the tolerance/resistance identified in this set of Solanum (Lycopersicon) accessions.     

<Emphasis Type="Italic">In vivo</Emphasis> Acclimatization Responses of <Emphasis Type="Italic">Platycodon grandiflorum</Emphasis> For. Duplex to Different Soil Types and Environmental Factors

This study aimed to evaluate the effects of soil types and environmental factors for optimum conditions of seedlings growth of the Platycodon grandiflorum for establishing the in vivo acclimatization system of regenerated plants derived from the in vitro culture. P. grandflorum seedlings were transferred to the in vivo condition and acclimatized under different soil types, light intensities, and various temperatures. Changes caused by environmental factors and soil types in plant growth viz. plant height, leaf width, leaf length, stem diameter, number of leaves, branches and nodes were recorded in this study. Among the nine types of soil, the best growth performances were obtained from the soil type SVP (Soil mixed with horticultural bed soil, vermiculite, and perlite @ 2:1:1). Seedlings of P. grandiflorum showed the best growth at higher levels of light intensity (60 μmol·m^-2·s^-1). In contrast, P. grandiflorum seedlings showed the best growth response at a moderate level of temperature (25°C). Collectively, the present study provides a better understanding of the responses of growth characteristics in P. grandiflorum seedlings exposed to various soil types, light intensities, and temperatures.