Mapping QTLs using a novel source of salinity tolerance from Hasawi and their interaction with environments in rice

Background

Salinity is one of the most severe and widespread abiotic stresses that affect rice production. The identification of major-effect quantitative trait loci (QTLs) for traits related to salinity tolerance and understanding of QTL × environment interactions (QEIs) can help in more precise and faster development of salinity-tolerant rice varieties through marker-assisted breeding. Recombinant inbred lines (RILs) derived from IR29/Hasawi (a novel source of salinity) were screened for salinity tolerance in the IRRI phytotron in the Philippines (E1) and in two other diverse environments in Senegal (E2) and Tanzania (E3). QTLs were mapped for traits related to salinity tolerance at the seedling stage.

Results

The RILs were genotyped using 194 polymorphic SNPs (single nucleotide polymorphisms). After removing segregation distortion markers (SDM), a total of 145 and 135 SNPs were used to construct a genetic linkage map with a length of 1655 and 1662 cM, with an average marker density of 11.4 cM in E1 and 12.3 cM in E2 and E3, respectively. A total of 34 QTLs were identified on 10 chromosomes for five traits using ICIM-ADD and segregation distortion locus (SDL) mapping (IM-ADD) under salinity stress across environments. Eight major genomic regions on chromosome 1 between 170 and 175 cM (qSES1.3, qSES1.4, qSL1.2, qSL1.3, qRL1.1, qRL1.2, qFWsht1.2, qDWsht1.2), chromosome 4 at 32 cM (qSES4.1, qFWsht4.2, qDWsht4.2), chromosome 6 at 115 cM (qFWsht6.1, qDWsht6.1), chromosome 8 at 105 cM (qFWsht8.1, qDWsht8.1), and chromosome 12 at 78 cM (qFWsht12.1, qDWsht12.1) have co-localized QTLs for the multiple traits that might be governing seedling stage salinity tolerance through multiple traits in different phenotyping environments, thus suggesting these as hot spots for tolerance of salinity. Forty-nine and 30 significant pair-wise epistatic interactions were detected between QTL-linked and QTL-unlinked regions using single-environment and multi-environment analyses.

Conclusions

The identification of genomic regions for salinity tolerance in the RILs showed that Hasawi possesses alleles that are novel for salinity tolerance. The common regions for the multiple QTLs across environments as co-localized regions on chromosomes 1, 4, 6, 8, and 12 could be due to linkage or pleiotropic effect, which might be helpful for multiple QTL introgression for marker-assisted breeding programs to improve the salinity tolerance of adaptive and popular but otherwise salinity-sensitive rice varieties.

     

Large-scale deployment of a rice 6 K SNP array for genetics and breeding applications

Background

Fixed arrays of single nucleotide polymorphism (SNP) markers have advantages over reduced representation sequencing in their ease of data analysis, consistently higher call rates, and rapid turnaround times. A 6 K SNP array represents a cost-benefit “sweet spot” for routine genetics and breeding applications in rice. Selection of informative SNPs across species and subpopulations during chip design is essential to obtain useful polymorphism rates for target germplasm groups. This paper summarizes results from large-scale deployment of an Illumina 6 K SNP array for rice.

Results

Design of the Illumina Infinium 6 K SNP chip for rice, referred to as the Cornell_6K_Array_Infinium_Rice (C6AIR), includes 4429 SNPs from re-sequencing data and 1571 SNP markers from previous BeadXpress 384-SNP sets, selected based on polymorphism rate and allele frequency within and between target germplasm groups. Of the 6000 attempted bead types, 5274 passed Illumina’s production quality control. The C6AIR was widely deployed at the International Rice Research Institute (IRRI) for genetic diversity analysis, QTL mapping, and tracking introgressions and was intensively used at Cornell University for QTL analysis and developing libraries of interspecific chromosome segment substitution lines (CSSLs) between O. sativa and diverse accessions of O. rufipogon or O. meridionalis. Collectively, the array was used to genotype over 40,000 rice samples. A set of 4606 SNP markers was used to provide high quality data for O. sativa germplasm, while a slightly expanded set of 4940 SNPs was used for O. sativa X O. rufipogon populations. Biparental polymorphism rates were generally between 1900 and 2500 well-distributed SNP markers for indica x japonica or interspecific populations and between 1300 and 1500 markers for crosses within indica, while polymorphism rates were lower for pairwise crosses within U.S. tropical japonica germplasm. Recently, a second-generation array containing ~7000 SNP markers, referred to as the C7AIR, was designed by removing poor-performing SNPs from the C6AIR and adding markers selected to increase the utility of the array for elite tropical japonica material.

Conclusions

The C6AIR has been successfully used to generate rapid and high-quality genotype data for diverse genetics and breeding applications in rice, and provides the basis for an optimized design in the C7AIR.

     

Frequency and fertility restoration efficiency of Rf3 and Rf4 genes in Indian rice

Identification of new parental lines is crucial for developing ecology‐specific hybrids with ideal agronomic performance. We screened a total of 570 different ecology‐specific Indian rice varieties for the presence of fertility restorer genes, Rf3 and Rf4 using tightly linked markers DRRM Rf3‐10 and RM6100, respectively. Among these varieties, 13% carried Rf3Rf3/Rf4Rf4, 31% carried rf3rf3/rf4rf4, 6% carried Rf3Rf3/rf4rf4 and remaining 50% carried Rf4Rf4/rf3rf3 allelic combinations. A mini set of 40 varieties with variable allelic combinations of fertility restorer genes were testcrossed with WA and Kalinga‐based CMS lines. All the 80 F₁s were evaluated for spikelet fertility and fertility restoration ability. Rf3Rf3/rf4rf4 genotypes mostly behaved as partial maintainers or partial restorers. In contrast, rf3rf3/Rf4Rf4 genotypes were partial or effective restorers. However, double dominant genotypes showed better fertility restoration than the genotypes containing Rf3 or Rf4 individually. Some of the genotypes showed unexpected restoration pattern implying occurrence of other fertility restorer(s) apart from Rf3 and Rf4. The perfect restorers and maintainers identified in this study can be directly used in hybrid rice breeding.     

Alteration in important quality traits and antioxidant activities in Brassica oleracea with Ogura cybrid cytoplasm

In vegetable brassicas, cytoplasmic male sterility (CMS) system is the most preferred mechanism for hybrid seed production and Ogura cybrid cytoplasm is the only source used widely. Effects of the alien cybrid cytoplasm on important quality traits need to be understood for their effective use in breeding programme. In analysing 38 Ogura cybrid cytoplasm‐based cauliflower CMS lines, it was revealed that introgression of Ogura cytoplasm resulted in significant alteration in important quality traits. Cupric reducing antioxidant activity and ferric reducing ability of plasma values were increased up to 5–10 times in different genotypes. Among 38 analysed CMS lines, concentration of plant pigments such as anthocyanin, total chlorophylls and ascorbic acid was reduced in 21, 17 and 32 genotypes, respectively. However, the concentration of total carotenoids and β‐carotene was elevated in most of the CMS lines and total carotenoid was increased up to 20 times in the CMS line, Ogu13‐85‐2A. The results indicate the role of nuclear–cytoplasmic interaction and mitochondrial genome in determining concentration of different quality traits.     

Extension of shelf life of tuberose flowers using a combination of gamma irradiation and generally regarded as safe (GRAS) preservatives and assessment of antimicrobial potency of senesced flowers

Tuberose cut flowers, available as loose flowers, were treated with gamma (γ) irradiation and generally regarded as safe (GRAS) preservative solutions for extension of shelf life. The flowers were packaged in low-density polyethylene bags, heat sealed and stored at 23 ± 2°C, 80% relative humidity (RH) and 4 ± 1°C, 40% RH, respectively. The flowers stored at these two temperature regimes were subjected to sensory evaluation and biochemical analyses. From these assessments, the longest shelf life of tuberose flowers was found to be 8 days at 23 ± 2°C, 80% RH (compared to 4 days for control) and 24 days at 4 ± 1°C, 40% RH (compared to 8 days for control) using combination treatment of low dose γ-irradiation (0.02 kGy) and preservative solutions (4% sucrose and 0.02% CaCl₂). Ethanolic extract of tuberose flowers of the most shelf stable set (stored at 4 ± 1°C), i.e. at the end of 24 days, showed antimicrobial potency against the common skin pathogen Staphylococcus aureus (ATCC 25923 and MDR strains), suggesting utility of the senesced tuberose flowers for therapeutic applications. This preservation technique would promote export of tuberose flowers by extension of their shelf lives and allow utilization of these flowers, post senescence.     

Comparisons of energy balance and evapotranspiration between flooded and aerobic rice fields in the Philippines

The seasonal and annual variability of sensible heat flux (H), latent heat flux (LE), evapotranspiration (ET), crop coefficient (K_c) and crop water productivity (WP_ET) were investigated under two different rice environments, flooded and aerobic soil conditions, using the eddy covariance (EC) technique during 2008-2009 cropping periods. Since we had only one EC system for monitoring two rice environments, we had to move the system from one location to the other every week. In total, we had to gap-fill an average of 50-60% of the missing weekly data as well as those values rejected by the quality control tests in each rice field in all four cropping seasons. Although the EC method provides a direct measurement of LE, which is the energy used for ET, we needed to correct the values of H and LE to close the energy balance using the Bowen ratio closure method before we used LE to estimate ET. On average, the energy balance closure before correction was 0.72 ± 0.06 and it increased to 0.99 ± 0.01 after correction. The G in both flooded and aerobic fields was very low. Likewise, the energy involved in miscellaneous processes such as photosynthesis, respiration and heat storage in the rice canopy was not taken into consideration.Average for four cropping seasons, flooded rice fields had 19% more LE than aerobic fields whereas aerobic rice fields had 45% more H than flooded fields. This resulted in a lower Bowen ratio in flooded fields (0.14 ± 0.03) than in aerobic fields (0.24 ± 0.01). For our study sites, evapotranspiration was primarily controlled by net radiation. The aerobic rice fields had lower growing season ET rates (3.81 ± 0.21 mm d⁻¹) than the flooded rice fields (4.29 ± 0.23 mm d⁻¹), most probably due to the absence of ponded water and lower leaf area index of aerobic rice. Likewise, the crop coefficient, K_c, of aerobic rice was significantly lower than that of flooded rice. For aerobic rice, K_c values were 0.95 ± 0.01 for the vegetative stage, 1.00 ± 0.01 for the reproductive stage, 0.97 ± 0.04 for the ripening stage and 0.88 ± 0.03 for the fallow period, whereas, for flooded rice, K_c values were 1.04 ± 0.04 for the vegetative stage, 1.11 ± 0.05 for the reproductive stage, 1.04 ± 0.05 for the ripening stage and 0.93 ± 0.06 for the fallow period. The average annual ET was 1301 mm for aerobic rice and 1440 mm for flooded rice. This corresponds to about 11% lower total evapotranspiration in aerobic fields than in flooded fields. However, the crop water productivity (WP_ET) of aerobic rice (0.42 ± 0.03 g grain kg⁻¹ water) was significantly lower than that of flooded rice (1.26 ± 0.26 g grain kg⁻¹ water) because the grain yields of aerobic rice were very low since they were subjected to water stress.The results of this investigation showed significant differences in energy balance and evapotranspiration between flooded and aerobic rice ecosystems. Aerobic rice is one of the promising water-saving technologies being developed to lower the water requirements of the rice crop to address the issues of water scarcity. This information should be taken into consideration in evaluating alternative water-saving technologies for environmentally sustainable rice production systems.     

Earthworm biomass response to soil management in semi-arid tropical Alfisol agroecosystems

Earthworms recorded during 1989–1993 across 15 soil management treatments, comprising three different tillagexthree organic amendments (bare, farmyard manure, and rice straw) and six perennial ley treatments, belonged to two endogeic species, Octochaetona phillotti (Michaelsen) and Lampito mauritii Kinberg, while in a nearby undisturbed natural revegetation area three species were found, including the above two and Octonochaeta rosea (Stephenson). The earthworm biomass showed significant temporal and spatial variations and was higher during the post monsoon period than in the early rainy season. No worm biomass was recorded during the dry season. In the tillage and organic amendment treatments, the biomass was drastically reduced from September 1989 to September 1991 after the application of carbofuran and some herbicides, and was significantly reduced during these two years compared to that of 1992. The maximum monthly earthworm biomass ranged between 2.5 and 17.9 g m^-2 across the treatments and increased several-fold in the nearby natural revegetation area (75.9 g m^-2). It significantly increased in perennial ley treatments compared to annual treatments with organic amendments. Thus the earthworm biomass varied significantly (P<0.01) across the 15 treatments, indicating discernible effects of soil management.Visiting Scientist (under the Rockefeller Foundation Environmental Research Fellowship in International Agriculture)     

Methane emission from two Indian soils planted with different rice cultivars

In a greenhouse study, methane emissions were measured from two diverse Indian rice-growing soils planted to five rice cultivars under similar water regimes, fertilizer applications and environmental conditions. Significant variations were observed in methane emitted from soils growing different cultivars. Total methane emission varied between 8.04 and 20.92gm^–2 from IARI soil (Inceptisol) and between 1.47 and 10.91gm^–2 from Raipur soil (Vertisol) planted to rice. In all the cultivars, emissions from IARI soil were higher than from Raipur soil. The first methane flux peak was noticed during the reproductive phase and the second peak coincided with the grain-ripening stage of the rice cultivars. Received: July 7, 1996     

Heavy Metal Contamination of Soil and Vegetation in the Vicinity of Industries in Bangladesh

Water, Air, &; Soil Pollution - A study was conducted to investigate the heavy metal contamination of soil and vegetation in the vicinity of industries around Dhaka city in Bangladesh....