Multi-environment evaluation of rice genotypes: impact of weather and culm biochemical parameters against sheath blight infection.

Introduction: Sheath blight caused by Rhizoctonia solani is one of the major diseases of rice, causing widespread crop losses. The use of semi-dwarf rice varieties in the ongoing nutrient-intensive rice cultivation system has further accentuated the incidence of the disease. An ideal solution to this problem would be identifying a stable sheath blight-tolerant genotype. Material and methods: A multi-environment evaluation of 32 rice genotypes against sheath blight infection was conducted over six seasons across two locations (Agricultural Research Farm, Institute of Agricultural Sciences, Banaras Hindu University (28.18° N, 38.03° E, and 75.5 masl), for four years during the wet seasons (kharif) from 2015 to 2018 and two seasons at the National Rice Research Institute (20°27'09" N, 85°55'57" E, 26 masl), Cuttack, Odisha, during the dry season (rabi) of 2019 and the kharif of 2019, including susceptible and resistant check. Percent disease index data were collected over 4 weeks (on the 7th, 14th, 21st, and 28th day after infection), along with data on other morphological and physiological traits. Result and discussion: The resistant genotypes across seasons were the ones with a higher hemicellulose content (13.93-14.64) and lower nitrogen content (1.10- 1.31) compared with the susceptible check Tapaswini (G32) (hemicellulose 12.96, nitrogen 1.38), which might explain the resistant reaction. Three different stability models-additive main effect and multiplicative interaction (AMMI), genotype + genotype x environment (GGE) biplot, and multi-trait stability index (MTSI)-were then used to identify the stable resistant genotypes across six seasons. The results obtained with all three models had common genotypes highlighted as stable and having a low area under the disease progress curve (AUDPC) values. The ideal stable genotypes with low disease incidence were IC 283139 (G19), Tetep (G28), IC 260917 (G4), and IC 277274 (G10), with AUDPC values of 658.91, 607.46, 479.69, and 547.94, respectively. Weather parameters such as temperature, rainfall, sunshine hours, and relative humidity were also noted daily. Relative humidity was positively correlated with the percent disease index.

Gene based markers improve precision of genome-wide association studies and accuracy of genomic predictions in rice breeding.

It is hypothesized that the genome-wide genic markers may increase the prediction accuracy of genomic selection for quantitative traits. To test this hypothesis, a set of candidate gene-based markers for yield and grain traits-related genes cloned across the rice genome were custom-designed. A multi-model, multi-locus genome-wide association study (GWAS) was performed using new genic markers developed to test their effectiveness for gene discovery. Two multi-locus models, FarmCPU and mrMLM, along with a single-locus mixed linear model (MLM), identified 28 significant marker-trait associations. These associations revealed novel causative alleles for grain weight and pleiotropic associations with other traits. For instance, the marker YD91 derived from the gene OsAAP3 on chromosome 1 was consistently associated with grain weight, while the gene has a significant effect on grain yield. Furthermore, nine genomic selection methods, including regression-based and machine learning-based models, were used to predict grain weight using a leave-one-out five-fold cross-validation approach to optimize the genomic selection model with genic markers. Among nine prediction models, Kernel Hilbert Space Regression (RKHS) is the best among regression-based models, and Random Forest Regression (RFR) is the best among machine learning-based models. Genomic prediction accuracies with and without GWAS significant markers were compared to assess the effectiveness of markers. The rapid decreases in prediction accuracy upon dropping GWAS significant markers indicate the effectiveness of new genic markers in genomic selection. Apart from that, the candidate gene-based markers were found to be more effective in genomic selection programs for better accuracy.

cgSSR marker-based genome-wide association study identified genomic regions for panicle characters and yield in rice (Oryza sativa L.).

BACKGROUND: To improve production efficiency, positive alleles corresponding to yield-related attributes must be accumulated in a single elite background. We designed and used cgSSR markers, which are superior to random SSR markers in genome-wide association study, to identify genomic regions that contribute to panicle characters and grain yield in this study. RESULTS: As evidenced by the high polymorphic information content value and gene diversity coefficient, the new cgSSR markers were determined to be highly informative. These cgSSR markers were employed to generate genotype data for an association panel evaluated for four panicle characters and grain yield over three seasons. For five traits, 17 significant marker-trait associations on six chromosomes were discovered. The percentage of phenotypic variance that could be explained ranged from 4% to 13%. Unrelated gene-derived markers had a strong association with target traits as well. CONCLUSION: Trait-associated cgSSR markers derived from corresponding or related genes ensure their utility in direct allele selection, while other linked markers aid in allele selection indirectly by altering the phenotype of interest. Through a marker-assisted breeding approach, these marker-trait associations can be leveraged to accumulate favourable alleles for yield enhancement in rice. © 2022 Society of Chemical Industry.

Recent developments in multi-omics and breeding strategies for abiotic stress tolerance in maize (Zea mays L.).

High-throughput sequencing technologies (HSTs) have revolutionized crop breeding. The advent of these technologies has enabled the identification of beneficial quantitative trait loci (QTL), genes, and alleles for crop improvement. Climate change have made a significant effect on the global maize yield. To date, the well-known omic approaches such as genomics, transcriptomics, proteomics, and metabolomics are being incorporated in maize breeding studies. These approaches have identified novel biological markers that are being utilized for maize improvement against various abiotic stresses. This review discusses the current information on the morpho-physiological and molecular mechanism of abiotic stress tolerance in maize. The utilization of omics approaches to improve abiotic stress tolerance in maize is highlighted. As compared to single approach, the integration of multi-omics offers a great potential in addressing the challenges of abiotic stresses of maize productivity.

Identification of microRNAs That Provide a Low Light Stress Tolerance-Mediated Signaling Pathway during Vegetative Growth in Rice.

Low light intensity affects several physiological parameters during the different growth stages in rice. Plants have various regulatory mechanisms to cope with stresses. One of them is the differential and temporal expression of genes, which is governed by post-transcriptional gene expression regulation through endogenous miRNAs. To decipher low light stress-responsive miRNAs in rice, miRNA expression profiling was carried out using next-generation sequencing of low-light-tolerant (Swarnaprabha) and -sensitive (IR8) rice genotypes through Illumina sequencing. Swarnaprabha and IR8 were subjected to 25% low light treatment for one day, three days, and five days at the active tillering stage. More than 43 million raw reads and 9 million clean reads were identified in Swarnaprabha, while more than 41 million raw reads and 8.5 million clean reads were identified in IR8 after NGS. Importantly, 513 new miRNAs in rice were identified, whose targets were mostly regulated by the genes involved in photosynthesis and metabolic pathways. Additionally, 114 known miRNAs were also identified. Five novel (osa-novmiR1, osa-novmiR2, osa-novmiR3, osa-novmiR4, and osa-novmiR5) and three known (osa-miR166c-3p, osa-miR2102-3p, and osa-miR530-3p) miRNAs were selected for their expression validation through miRNA-specific qRT-PCR. The expression analyses of most of the predicted targets of corresponding miRNAs show negative regulation. Hence, miRNAs modulated the expression of genes providing tolerance/susceptibility to low light stress. This information might be useful in the improvement of crop productivity under low light stress.

Understanding natural genetic variation for grain phytic acid content and functional marker development for phytic acid-related genes in rice.

BACKGROUND: The nutritional value of rice can be improved by developing varieties with optimum levels of grain phytic acid (PA). Artificial low-PA mutants with impaired PA biosynthesis have been developed in rice through induced mutagenesis. However, low-PA mutant stocks with drastically reduced grain PA content have poor breeding potential, and their use in rice breeding is restricted due to their detrimental pleiotropic effects, which include decreased seed viability, low grain weight, and low seed yield. Therefore, it is necessary to take advantage of the natural variation in grain PA content in order to reduce the PA content to an ideal level without compromising the crop's agronomic performance. Natural genetic diversity in grain PA content has not been thoroughly examined among elite genetic stocks. Additionally, given grain PA content as a quantitative trait driven by polygenes, DNA marker-assisted selection may be required for manipulation of such a trait; however, informative DNA markers for PA content have not yet been identified in rice. Here we investigated and dissected natural genetic variation and genetic variability components for grain PA content in rice varieties cultivated in Eastern and North-Eastern India during the last 50 years. We developed novel gene-based markers for the low-PA-related candidate genes in rice germplasm, and their allelic diversity and association with natural variation in grain PA content were studied. RESULTS: A wide (0.3-2.8%), significant variation for grain PA content, with decade-wise and ecology-wise differences, was observed among rice varieties. Significant genotype x environment interaction suggested polygenic inheritance. The novel candidate gene-based markers detected 43 alleles in the rice varieties. The new markers were found highly informative as indicated by PIC values (0.11-0.65; average: 0.34) and coverage of total diversity. Marker alleles developed from two putative transporter genes viz., SPDT and OsPT8 were significantly associated with grain PA variation assayed on the panel. A 201 bp allele at the 3' UTR of SPDT gene was negatively associated with grain PA content and explained 7.84% of the phenotypic variation. A rare allele in the coding sequence of OsPT8 gene was positively associated with grain PA content which explained phenotypic variation of 18.49%. CONCLUSION: Natural variation in grain PA content is substantial and is mostly controlled by genetic factors. The unique DNA markers linked with PA content have significant potential as genomic resources for the development of low-PA rice varieties through genomics-assisted breeding procedures.

Unravelling genetic architecture and development of core set from elite rice lines using yield-related candidate gene markers.

Assessing genetic diversity and development of a core set of elite breeding lines is a prerequisite for selective hybridization programes intended to improve the yield potential in rice. In the present study, the genetic diversity of newly developed elite lines derived from indicax tropical japonica and indicax indica crosses were estimated by 38 reported molecular markers. The markers used in the study consist of 24 gene-based and 14 random markers related to grain yield-related QTLs distributed across the rice genome. Genotypic characterization was carried out to determine the genetic similarities between the elite lines. In total, 75 alleles were found using 38 polymorphic markers, with polymorphism information content ranging from 0.10 to 0.51 with an average of 0.35. The genotypes were divided into three groups based on cluster analysis, structure analysis and also dispersed throughout the quadrangle of PCA, but nitrogen responsive lines clustered in one quadrangle. Seven markers (GS3_RGS1, GS3_RGS2, GS5_Indel1, Ghd 7_05SNP, RM 12289, RM 23065 and RM 25457) exhibited PIC values ≥ 0.50 indicating that they were effective in detecting genetic relationships among elite rice. Additionally, a core set of 11 elite lines was made from 96 lines in order to downsize the diversity of the original population into a small set for parental selection. In general, the genetic information collected in this work will aid in the study of grain yield traits at molecular level for other sets of rice genotypes and for selecting diverse elite lines to develop a strong crossing programme in rice. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01190-8.

Understanding complex genetic architecture of rice grain weight through QTL-meta analysis and candidate gene identification.

Quantitative trait loci (QTL) for rice grain weight identified using bi-parental populations in various environments were found inconsistent and have a modest role in marker assisted breeding and map-based cloning programs. Thus, the identification of a consistent consensus QTL region across populations is critical to deploy in marker aided breeding programs. Using the QTL meta-analysis technique, we collated rice grain weight QTL information from numerous studies done across populations and in diverse environments to find constitutive QTL for grain weight. Using information from 114 original QTL in meta-analysis, we discovered three significant Meta-QTL (MQTL) for grain weight on chromosome 3. According to gene ontology, these three MQTL have 179 genes, 25 of which have roles in developmental functions. Amino acid sequence BLAST of these genes indicated their orthologue conservation among core cereals with similar functions. MQTL3.1 includes the OsAPX1, PDIL, SAUR, and OsASN1 genes, which are involved in grain development and have been discovered to play a key role in asparagine biosynthesis and metabolism, which is crucial for source-sink regulation. Five potential candidate genes were identified and their expression analysis indicated a significant role in early grain development. The gene sequence information retrieved from the 3 K rice genome project revealed the deletion of six bases coding for serine and alanine in the last exon of OsASN1 led to an interruption in the synthesis of α-helix of the protein, which negatively affected the asparagine biosynthesis pathway in the low grain weight genotypes. Further, the MQTL3.1 was validated using linked marker RM7197 on a set of genotypes with extreme phenotypes. MQTL that have been identified and validated in our study have significant scope in MAS breeding and map-based cloning programs for improving rice grain weight.

Identification of QTLs for zinc deficiency tolerance in a recombinant inbred population of rice (Oryza sativa L.).

BACKGROUND: Deficiency of Zn is a major soil constraint in rice plant growth and yield. Edaphic factors such as Zn deficiency in soil in relation to plant performance are still poorly understood. Here, we report promising quantitative trait loci (QTL) conferring tolerance to Zn deficiency, which were identified through biparental mapping. The experiment was conducted using the 236 F7 recombinant inbred line mapping population derived from the cross of Kinandang Patong (Zn deficiency sensitive) and A69-1 (Zn deficiency tolerant). RESULTS: A total of six QTLs (qLB-2B, qLB-4B, qPM-4B, qPM-6B, qRZC-4B, qSZC-4B) on chromosomes 2, 4 and 6 were identified for environment 1, whereas five QTLs (qLB-2 N, qLB-4 N, qPM-4 N, qRZC-4 N, qSZC-4 N) on chromosomes 2 and 4 were detected for environment 2. Among these, five major (51.30, 48.70, 28.60, 56.00, 52.00 > 10 R2 ) and one minor (5.40 < 10 R2 ) QTLs for environment 1 and four major (51.48, 50.20, 53.00, 48.00 > 10 R2 ) and one minor (4.44 < 10) QTLs for environment 2 for Zn deficiency tolerance with a logarithm of odd threshold value higher than 3 were identified. The QTLs (qLB-4B, qPM-4B, qRZC-4B, qSZC-4B, qLB-4 N, qPM-4 N, qRZC-4 N, qSZC-4 N) for leaf bronzing, plant mortality root zinc concentration and shoot zinc concentration identified on chromosome 4 were found to be the most promising and highly reproducible across the locations that explained phenotypic variation from 48.00% to 56.00% with the same marker interval RM6748-RM303. CONCLUSION: The new QTLs and its linked markers identified in the present study can be utilized for Zn deficiency tolerance in elite cultivars using marker-assisted backcrossing. © 2022 Society of Chemical Industry.

The role of phytochrome-mediated gibberellic acid signaling in the modulation of seed germination under low light stress in rice (O. sativa L.).

Seed germination plays cardinal roles in seedling establishment and their successive growth. However, seed germination is retarded by far-red (FR) enrichment under low light stress, and the inhibitory signalling mechanism remains ambiguous. Our results indicated that low light treatment, both in the open and growth chamber conditions, inhibits rice seed germination by decreasing the gibberellin (GA) contents. To explore the mechanism of GA-deficiency under low light stress, differential expression profiling of GA-anabolic, -catabolic, ABA -anabolic, -catabolic, and SLR1 was investigated, revealing that expression of ABA- anabolic, GA-catabolic genes and SLR1 was upregulated with a simultaneous downregulation of ABA-catabolic and GA-anabolic genes under low light treatment. These results suggested that FR-induced GA inadequacy is resulted by upregulation of SLR1 and GA-catabolism genes consequently increase DELLA that further subsided GA-responses in the germinating rice seeds. Moreover, we provided evidence that FR-induced GA inadequacy demotes rice seed germination by decreasing amylase activity, eventually decreasing the carbohydrate solubilization in the germinating seeds. Finally, we suggest that under low light stress, due to a retarded conversion of phytochrome A to their bioactive form, the ABA-catabolic genes were eventually upregulated with a simultaneous downregulation of GA-anabolic genes. Consequently, a lower GA pool fails to leverage the GA-dependent DELLA degradation, further shutting down the expected GA responses that reduce germination efficiency under FR-enriched light. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01167-7.

A single nucleotide substitution in the SPDT transporter gene reduced phytic acid and increased mineral bioavailability from Rice grain (Oryza sativa L.).

Phosphorus (P) flow in agricultural land depends on the P taken off from harvested product, its losses through runoff and fertilizer applied to balance the removed P. Phytic acid (PA), the major storage form of phosphorus (P) in cereal grains is a key anti-nutrient for human and non-ruminants leads to eutrophication of waterways. As the natural non-renewable P reserves are limited, enhancing P use efficiency is needed for field crops. SULTR-like phosphorus distribution transporter (SPDT) is a novel rice transporter transfer P to the grain. Any alteration in transporter gene reduce grain P with concomitant rise in the leaves. A low PA (3.0 g/kg) rice Khira was identified where a single nucleotide mutation in LOC_Os06g05160 gene encoding SPDT showed low P transportation to grain. An amino acid change was detected as Valine-330 to Alanine at the 3' end of fifth exon. Highest expression of SPDT was observed in node I of rice as compared to low PA genotype. The mutation in SPDT could significantly affect P and PA accumulation in the grains with increased mineral bioavailability. PRACTICAL APPLICATIONS: Excessive P application in crop leads to higher production cost as well as rapid depletion of limited rock phosphate. Alteration of P transporter function in the rice lower PA and total P accumulation in the grains with increased mineral bioavailability. The re-distributed P in the straw can be applied as manure to the rice field. Thus, less P will be removed from the field, result in the decreased requirement for P fertilizer.

Non-destructive phenotyping for early seedling vigor in direct-seeded rice.

BACKGROUND: Early seedling vigor is an essential trait of direct-seeded rice. It helps the seedlings to compete with weeds for water and nutrient availability, and contributes to better seedling establishment during the initial phase of crop growth. Seedling vigor is a complex trait, and phenotyping by a destructive method limits the improvement of this trait through traditional breeding. Hence, a non-invasive, rapid, and precise image-based phenotyping technique is developed to increase the possibility to improve early seedling vigor through breeding in rice and other field crops. RESULTS: To establish and assess the methodology using free-source software, early seedling vigor was estimated from images captured with a digital SLR camera in a non-destructive way. Here, the legitimacy and strength of the method have been proved through screening seven diverse rice cultivars varying for early seedling vigor. In the regression analysis, whole-plant area (WPA) estimated by destructive-flatbed scanner (WPAs) and non-destructive imaging (WPAi) approaches was strongly related (R2 > 83%) and suggested that WPAi can be adapted in place of destructive methods to estimate seedling vigor. In addition, this study has identified a set of new geometric traits (convex hull and top view area) for screening breeding lines for early seedling vigor in rice, which decreased the time by 80% and halved the cost of labor in data observation. CONCLUSIONS: The method demonstrated here is affordable and easy to establish as a phenotypic platform. It is suitable for most glasshouses/net houses for characterizing genotypes to understand the plasticity of shoots under a given environment at the seedling stage. The methodology explained in this experiment has been proven to be practical and suggested as a technique for researchers involved in direct-seeded rice. Consequently, it will help in the simultaneous screening of genotypes in large numbers, the identification of donors, and in gaining information on the genetic basis of the trait to design a breeding program for direct-seeded rice.

Role of sedoheptulose-1,7 bisphosphatase in low light tolerance of rice (Oryza sativa L.).

Rice grain yield is drastically reduced under low light especially in kharif (wet) season due to cloudy weather during most part of crop growth. Therefore, 50-60% of yield penalty was observed. To overcome this problem, identification of low light tolerant rice genotypes with a high buffering capacity trait such as photosynthetic rate has to be developed. Sedoheptulose-1,7 bisphosphatase, a light-regulated enzyme, plays pivotal role in the Calvin cycle by regenerating the substrate (RuBP) for RuBisCo and therefore, indirectly regulates the influx of CO2 for this crucial process. We found a potential role of SBPase expression and activity in low light tolerant and susceptible rice genotypes by analyzing its influence on net photosynthetic rate and biomass. We observed a significant relationship of yield with photosynthesis, SBPase expression and activity especially under low light conditions. Two tolerant and two susceptible rice genotypes were used for the present study. Tolerant genotypes exhibited significant but least reduction compared to susceptible genotypes in the expression and activity of SBPase, which was also manifested in its photosynthetic rate and finally in the grain yield under low light. However, susceptible genotypes showed significant reduction in SBPase activity along with photosynthesis and grain yield suggesting that tracking the expression and activity of SBPase could form a simple and reliable method to identify the low light tolerant rice cultivars. The data were analyzed using the Indostat 7.5, Tukey-Kramer method through Microsoft Excel 2019 and PAST4.0 software. The significant association of SBPase activity with the grain yield, net assimilation rate, electron transfer rate, biomass and grain weight were observed under low light stress. These traits should be considered while selecting and breeding for low light tolerant cultivars. Thus, SBPase plays a major role in the low light tolerance mechanism in rice.

Evaluation of variability and environmental stability of grain quality and agronomic parameters of pigmented rice (O. sativa L.).

Eleven pigmented rice genotypes were evaluated to estimate genetic parameters, heritability and association. The results indicated that, genotypic variation was high among the lines. The distinct seasonal effect on plant performance for antioxidant capacity, anthocyanin, flavonoids, head rice recovery and test weights was also observed. Wet season favoured the crop performance in all genotypes as compared to drought conditions. The differential accumulation of different quality traits such as AOA, anthocyanin content, flavonoids content, etc showed high heritability, which would be transfer to high yeilding popular rice cultivars through conventional or geneticaly modification techniques. The line Mamihunger was chosen as donor of the high-quality rice grain and Annapurna for high yield. Further, Mamihunger are foreseen to be good in nutritional quality and industry use.