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.

     

Genome-wide association study for spot blotch resistance in Afghan wheat germplasm

Spot blotch (SB), caused by Bipolaris sorokiniana, is a devastating disease of wheat globally, especially in South Asia and South America. Understanding the genetics of resistance to SB is important for developing breeding strategies to improve resistance. A panel of 301 genotypes from Afghanistan was phenotyped over two crop seasons using a mixture of virulent B. sorokiniana isolates and genotyped using DArTSeq to obtain genome-wide markers. Fifty genotypes (16.6%) showed disease scores less than the resistant control. Principal component analysis using the genotypic data clustered the genotypes into five different groups. Among models used for genome-wide association mapping, the multilocus mixed model, and fixed and random model circulating probability unification algorithms were most effective in identifying significant marker-trait associations (MTA). Twenty-five MTAs at p ≤ .001 were identified on chromosomes 1A, 1B, 1D, 2B, 2D, 3A, 3B, 4A, 5A, 5B, 6A, 7A, and 7D, indicating the quantitative nature of resistance to SB. Phenotypic variation explained by these markers ranged from 2.0% to 17.7%, and genomic regions on the chromosomes 1D, 2D, 3A, 3B, 4A, 5A, and 5B coincided with loci identified in previous studies. Three single nucleotide polymorphism (SNP) markers on chromosomes 1B (SNP 1113207) and 5A (SNPs 5411867 and 998276) were significant in both crop seasons as well as in the combined analysis across seasons. Marker 5411867 is close to Vrn-A1, shown to be associated with SB in previous studies. Furthermore, among known SB resistance genes, Sb2 on chromosome 5B was predicted to be significant in this panel.     

Biocontrol potential of Pseudomonas azotoformans,Serratia marcescens and Trichoderma virens against Fusarium wilt of Dalbergia sissoo

Wilt disease caused by Fusarium solani is a serious constraint to Dalbergia sissoo (shisham) plantations in northern India. In this study, the antagonistic potential of 40 bacterial isolates recovered from rhizophere soil of healthy shisham trees, and a well‐characterized Trichoderma species (Trichoderma virens) were tested for their possibility as biocontrol agents for F. solani. Two promising isolates (S1 and S15) were identified which inhibited pathogen growth, caused chitin degradation, produced siderophores and solubilized phosphate in vitro. Isolate S15 scored highest for hydrogen cyanide (HCN) production while isolate S1 was a non‐HCN producer. These two isolates were identified as Serratia marcescens (S1) and Pseudomonas azotoformans (S15) following sequence analysis of 16S rDNA. In dual culture assays, T. virens caused 80% inhibition of mycelial growth of the test fungus. The three selected antagonists when tested in planta in the glasshouse completely suppressed production of wilt symptoms on 12‐month‐old shisham plants. Further work is needed to ascertain the potential of these isolates to be used as biocontrol agents to manage shisham wilt under field conditions.     

Rice-residue biochar influences phosphorus availability in soil with contrasting P status

ABSTRACT

Thermo-chemical conversion of crop residues to produce biochar is an emerging strategy in the context of sustainable phosphorous (P) use and residue management. An incubation study for 90 d was conducted to investigate the effects of rice-residue biochar (0, 10, 20 and 40 g kg^?1) in combination with inorganic-P (KH₂PO₄) (0, 25 and 50 mg kg^?1) on phosphorous availability in medium- and high-P status soils. Increasing biochar addition rates alone or in combination with inorganic-P resulted in a significant increase in P pools, i.e. plant available P or Olsen-P (from 8 to 132 mg kg^?1 in medium-P and 15 to 160 mg kg^?1 in high-P soils), microbial biomass P and various mineral-bound inorganic-P fractions in the order (Ca-P > organic-P > Al-P > loosely held/soluble-P > Fe-P > reductant soluble-P). Further, lower phosphatase activity (19–50%) with increasing rates of biochar addition in both soils elucidates the ability of biochar to act as a long-term source of available P in the experimental soils. The results demonstrate that rice-residue biochar can directly or indirectly enhance the status of available P in soils and hence can be used as a beneficial amendment to meet the crop P demand.     

Grain yield,nitrogen use efficiency and tillage intensity in wheat as affected by precision nutrient management

ABSTRACT

The present studies were conducted to evaluate the effect of different nutrient management practices under two tillage options in wheat. The experiments were laid out in split-plot design with a combination of two varieties (WH 1105 and HD 2967) and two tillage options (Conventional and No tillage) in the main plot and six precision nutrient management practices [absolute control, site-specific nutrient management with Nutrient Expert for wheat (SSNM-NE)(170 kg nitrogen (N)/ha), SSNM NE+GreenSeeker (GS)(153/158 N kg/ha), N₁₂₀ (120 kg N/ha) before irrigation, N₁₂₀ after irrigation and N Rich (180 kg N/ha)] in subplot replicated thrice. The grain yield and quality characters in no tillage (NT) and conventional tillage (CT) were similar but agronomic efficiency was higher in NT. Both the varieties (WH 1105 and HD 2967) gave similar grain yield and quality. Wheat variety WH 1105 recorded significantly higher sodium dodecyl sulfate sedimentation (SDS) and gluten index. The treatment SSNM NE+GS had resulted in 107.1% higher grain yield than no nitrogen control but similar to enriched N plot (180 kg N/ha). The grain protein, SDS and gluten index in need-based nutrient management (SSNM+GS) treatment were found to be similar as recorded in SSNM-NE (170 kgN/ha) and N enriched plot (180 kg N ha^?1). The agronomic efficiency and recovery efficiency in SSNM+GS were also better than SSNM NE.     

Hydrus-1D model for simulating water flow through paddy soils under alternate wetting and drying irrigation practice

Paddy and Water Environment - Alternate wetting and drying (AWD) irrigation practice in paddy cultivation requires a safe soil moisture stress level in the root zone for which irrigation water...     

Pigeonpea improvement: An amalgam of breeding and genomic research

In the past five decades, constant research has been directed towards yield improvement in pigeonpea resulting in the deployment of several commercially acceptable cultivars in India. Though, the genesis of hybrid technology, the biggest breakthrough, enigma of stagnant productivity still remains unsolved. To sort this productivity disparity, genomic research along with conventional breeding was successfully initiated at ICRISAT. It endowed ample genomic resource providing insight in the pigeonpea genome combating production constraints in a precise and speedy manner. The availability of the draft genome sequence with a large‐scale marker resource, oriented the research towards trait mapping for flowering time, determinacy, fertility restoration, yield attributing traits and photo‐insensitivity. Defined core and mini‐core collection, still eased the pigeonpea breeding being accessible for existing genetic diversity and developing stress resistance. Modern genomic tools like next‐generation sequencing, genome‐wide selection helping in the appraisal of selection efficiency is leading towards next‐generation breeding, an awaited milestone in pigeonpea genetic enhancement. This paper emphasizes the ongoing genetic improvement in pigeonpea with an amalgam of conventional breeding as well as genomic research.     

Inheritance and molecular mapping of restorer‐of‐fertility (Rf) gene in A2 hybrid system in pigeonpea (Cajanus cajan)

Inheritance of fertility restorer gene in pigeonpea was studied using F₂ and BC₁F₁ populations derived from cross AL103A × IC245273. It was found to be controlled by single dominant gene. Out of 228 SSR primer pairs, 33 primer pairs showed parental polymorphism, while four primers were found polymorphic in bulk segregant analysis (BSA). These four primers viz., CcM 1615, CcM 0710, CcM 0765 and CcM 1522 were used for genotyping of F₂ population and were found to be placed at 3.1, 5.1, 28.1 and 45.8 cM, respectively. Two of them, CcM 1615 and CcM 0710, evinced clear and unambiguous bands for fertility restoration in F₂ population. The Rf gene was mapped on linkage group 6 between the SSR markers CcM 1615 and CcM 0710 with the distances of 3.1 and 5.1 cM, respectively. The accuracy of the CcM 1615 was validated in 18 restorers and six maintainer lines. The marker CcM 1615 amplified in majority of male restorer lines with a selection accuracy of 91.66%.     

Achyranthes aspera (Prickly chaff flower) leaves- and seeds-supplemented diets regulate growth,innate immunity,and oxidative stress in Aeromonas hydrophila-challenged Labeo rohita

ABSTRACT

The immunostimulatory and disease-resistance properties of Achyranthes aspera were evaluated in rohu (Labeo rohita) challenged with Aeromonas hydrophila. Experimental diets were enriched with leaves at 0.25% (D1) and 0.5% (D2) and seeds at 0.5% (D3); the control diet (D4) was without any enrichment. Rohu (2.02 ± 0.23 g) were cultured for 75 days and then challenged with bacteria. The highest average weight was observed in the D3 diet-fed fish. The cumulative mortality rates were 70%, 60%, 40%, and 30% in the D4, D1, D2, and D3 diets fed to rohu respectively. Enriched diets significantly increased myeloperoxidase, nitric oxide synthase, and serum lysozyme levels and decreased malondialdehyde and carbonyl protein content. Expressions of lysozyme C and lysozyme G were significantly (P < .05) higher in the D3 diet-fed fish. In the kidney, IL-1β and TLR 4 were up-regulated in enriched-diet-fed fish. Supplementation of seeds and leaves at 0.5% showed a positive impact in fish.     

Soil carbon change across ten New South Wales farms under different farm management regimes in Australia

This paper examines the potential influence of soil management and land use on soil carbon on cropping farms in New South Wales (NSW), Australia. Soil organic carbon (SOC) data from ten farms spatially distributed across NSW were examined on two occasions. Soil cores to a depth 0–30 cm were measured for SOC and, as expected, SOC in the A horizon (1.16%) was significantly (p < .001) greater than in the B horizon (0.74%) of all profiles. Analysis of the 2013 and 2015 SOC data indicated that in many ways, the results runs counter to other SOC studies in Australia. Importantly, the mean SOC concentration in these agricultural soils was significantly (p < .001) less under cropping (2013-1.05%, 2015-0.97%) than in native sites (2013-1.20%, 2015-1.16%). Out of the total of 35 sites sampled from 10 farms, SOC in 49% of sites did not change significantly over 2 years, in 17% it increased significantly, whereas in 34% it decreased. Further, a clear implication of drought on SOC was seen on sites that were uncropped based on a critical value for a 95% confidence interval (p < .05) and complemented by the significant correlation (p < .05) between average annual precipitation deficit (ANPD) and SOC across the state with R² = 0.39. The mean SOC was found to be directly proportional to standard deviation and standard error. In terms of spatial variability, the C0 (nugget) value was greatest for farms with a large mean SOC and the average variogram in this study has a range of approximately 200 m which is potentially useful in determining sampling spacing for soil carbon auditing purpose. Similar empirical data over more years are required to better estimate SOC levels and to determine whether at a farm scale, factors such as land management, land use and climate can be related to soil carbon change and variability.