Rice Chromosome Research Articles

Fine Mapping of Two Interacting Loci for Transmission Ratio Distortion in Rice (Oryza sativa L.)

Transmission ratio distortion (TRD) denotes the observed allelic or genotypic frequency deviation from the expected Mendelian segregation ratios in the offspring of a heterozygote. TRD can severely hamper gene flow between and within rice species. Here, we report the fine mapping and characterization of two loci (TRD4.1 and TRD4.2) for TRD using large F2 segregating populations, which are derived from rice chromosome segment substitution lines, each containing a particular genomic segment introduced from the japonica cultivar Nipponbare (NIP) into the indica cultivar Zhenshan (ZS97). The two loci exhibited a preferential transmission of ZS97 alleles in the derived progeny. Reciprocal crossing experiments using near-isogenic lines harboring three different alleles at TRD4.1 suggest that the gene causes male gametic selection. Moreover, the transmission bias of TRD4.2 was diminished in heterozygotes when they carried homozygous TRD4.1ZS97. This indicates an epistatic interaction between these two loci. TRD4.2 was mapped into a 35-kb region encompassing one candidate gene that is specifically expressed in the reproductive organs in rice. These findings broaden the understanding of the genetic mechanisms of TRD and offer an approach to overcome the barrier of gene flow between the subspecies in rice, thus facilitating rice improvement by introgression breeding.

Year-round flowering gene e1, a mutation at the E1 locus on rice chromosome 7 and its combination with green revolution gene sd1 in an isogenic cell line.

Genetic analysis on the year-round flowering gene e1, which was derived from Kanto No. 79, an induced mutant by Koshihikari gamma-ray irradiated, was conducted through the backcross process to combine e1 and sd1 in the genetic background of Koshihikari. e1 strongly forwarded flowering 14 days earlier than the original variety Koshihikairi. Isogenic Koshihiakri combining e1 and sd1 was developed by four times of backcross with either Koshihikari or Koshihikari sd1 as recurrent parents by using the e1sd1 homozygous F3 plant in Koshihikari sd1 × Kanto No. 79 as a non-recurrent parent. As a result, "Koshihikari e1sd1" maturing 14 days earlier was approximately 30 cm shorter than Koshihikari. e1 was linked with DNA markers, which were near to Ghd7 on the short arm of chromosome 7. The whole genome sequencing revealed a single candidate SNP, which is specific to Koshihikari e1sd1, in the Xa21-like sequence, at 35213 bp downstream from Ghd7 on chromosome 7. Koshihikari e1sd1-specific SNP beside Ghd7 is expected to downregulate with Ghd7.

Transcriptomic identification of long noncoding RNAs and their hormone-associated nearby coding genes involved in the differential development of caryopses localized on different branches in rice

Long noncoding RNAs (lncRNAs) play important regulatory roles in caryopsis development and grain size in rice. However, whether there exist differences in lncRNA expression between caryopses located on primary branches (CPB) and caryopses located on secondary branches (CSB) that contribute to their differential development remains elusive. Here, we performed transcriptome-wide analysis to identify 2,273 lncRNAs expressed in CPB and CSB at 0, 5, 12, and 20 days after flowering (DAF). Although these lncRNAs were widely distributed, the majority were located in intergenic regions of the 12 rice chromosomes. Based on gene expression cluster analysis, lncRNAs expressed in CPB and CSB were clustered into two subtypes in a position-independent manner: one includes 0- and 5-DAF CPB and CSB, and 12-DAF CSB; the second includes 12-DAF CPB and 20-DAF CPB and CSB. Furthermore, according to the expression value of each lncRNA, K-means cluster analysis revealed 135 early-stage, 116 middle-stage, and 114 late-stage expression-delayed lncRNAs in CSB. Then, we analyzed the expression values of the expression-delayed lncRNAs and nearby coding genes (100 kb upstream and downstream of the lncRNAs), and found 631 lncRNA–mRNA pairs, including 258 lncRNAs and 571 nearby coding genes, some of which are related to hormone-regulated grain development. These results suggested that expression-delayed lncRNAs in CSB may regulate the development of CPB and CSB, providing insight into the mechanism underlying the developmental differences between CPB and CSB, and the differences in grain yield.

Meta-QTLs linked to nitrogen use efficiency are randomly distributed in Indian rice germplasm

Nitrogen (N) recognized as a critical element for plant growth plays a fundamental role in rice cultivation. The N use efficiency (NUE) hovers around 30-35% in rice, suggesting a significant loss of N from the rice fields. Improving the NUE therefore would require genetic interventions and breeding. The cultivar improvement for N uptake and utilization is required to elevate NUE to further heights. Several quantitative trait loci (QTLs) for NUE under varying conditions and genetic backgrounds have been reported in rice. Consolidation of this distributed and unorganized information is necessary to identify critical genomic regions to be used for crop improvement. Therefore, a Meta-analysis from an assembly of 506 QTLs reported from 18 different studies was performed to identify the most significant genomic regions associated with NUE in rice. A total of 12 meta-QTLs (mQTLs) related to the traits such as NUE and grain yield per plant under N deficit conditions have been identified over four rice chromosomes namely 1, 3, 4, and 8. Evaluation of these mQTLs in a set of Indian rice germplasm revealed a significant association of the meta loci with N use parameters and showed wide distribution in the germplasm. Identification of mQTLs on different chromosomes together with their respective markers will help recruit them in marker-assisted selection (MAS) to develop N use efficient genotypes.

Genome‑wide identification of CAMTA gene family members in rice (Oryza sativa L.) and in silico study on their versatility in respect to gene expression and promoter structure

The calmodulin-binding transcription activator (CAMTA) is a family of transcriptional factors containing a cluster of calmodulin-binding proteins that can activate gene regulation in response to stresses. The presence of this family of genes has been reported earlier, though, the comprehensive analyses of rice CAMTA (OsCAMTA) genes, their promoter regions, and the proteins were not deliberated till date. The present report revealed the existence of seven CAMTA genes along with their alternate transcripts in five chromosomes of rice (Oryza sativa) genome. Phylogenetic trees classified seven CAMTA genes into three clades indicating the evolutionary conservation in gene structure and their association with other plant species. The in silico study was carried out considering 2 kilobases (kb) promoter regions of seven OsCAMTA genes regarding the distribution of transcription factor binding sites (TFbs) of major and plant-specific transcription factors whereas OsCAMTA7a was identified with highest number of TFbs, while OsCAMTA4 had the lowest. Comparative modelling, i.e., homology modelling, and molecular docking of the CAMTA proteins contributed the thoughtful comprehension of protein 3D structures and protein-protein interaction with probable partners. Gene ontology annotation identified the involvement of the proteins in biological processes, molecular functions, and localization in cellular components. Differential gene expression study gave an insight on functional multiplicity to showcase OsCAMTA3b as most upregulated stress-responsive gene. Summarization of the present findings can be interpreted that OsCAMTA gene duplication, variation in TFbs available in the promoters, and interactions of OsCAMTA proteins with their binding partners might be linked to tolerance against multiple biotic and abiotic cues.

Early Domestication History of Asian Rice Revealed by Mutations and Genome-Wide Analysis of Gene Genealogies

BackgroundAsian rice (Oryza sativa L.) has been a model plant but its cultivation history is inadequately understood, and its origin still under debate. Several enigmas remain, including how this annual crop shifted its growth habit from its perennial ancestor, O. rufipogon, why genetic divergence between indica and japonica appears older than the history of human domestication, and why some domestication genes do not show signals of introgression between subgroups. Addressing these issues may benefit both basic research and rice breeding.ResultsGene genealogy-based mutation (GGM) analysis shows that history of Asian rice is divided into two phases (Phase I and II) of about equal lengths. Mutations occurred earlier than the partition of indica and japonica to Os genome mark Phase-I period. We diagnosed 91 such mutations among 101 genes sampled across 12 chromosomes of Asian rice and its wild relatives. Positive selection, detected more at 5′ regions than at coding regions of some of the genes, involved 22 loci (e.g., An-1, SH4, Rc, Hd3a, GL3.2, OsMYB3, OsDFR, and OsMYB15), which affected traits from easy harvesting, grain color, flowering time, productivity, to likely taste and tolerance. Phase-I mutations of OsMYB3, OsHd3a and OsDFR were experimentally tested and all caused enhanced functions of the genes in vivo. Phase-II period features separate cultivations, lineage-specific selection, and expanded domestication to more genes. Further genomic analysis, along with phenotypic comparisons, indicates that O. sativa is hybrid progeny of O. rufipogon and O. nivara, inherited slightly more genes of O. rufipogon. Congruently, modern alleles of the sampled genes are approximately 6% ancient, 38% uni-specific, 40% bi-specific (mixed), and 15% new after accumulating significant mutations. Results of sequencing surveys across modern cultivars/landraces indicate locus-specific usages of various alleles while confirming the associated mutations.ConclusionsAsian rice was initially domesticated as one crop and later separate selection mediated by human resulted in its major subgroups. This history and the hybrid origin well explain previous puzzles. Positive selection, particularly in 5′ regions, was the major force underlying trait domestication. Locus-specific domestication can be characterized and the result may facilitate breeders in developing better rice varieties in future.

Parental Polymorphic Marker Survey and Genetic Diversity Studies among the Popular Maintainer Lines of Hybrid Rice (Oryza sativa L.) for Stigma Exsertion Trait

Parental polymorphic survey using rice satellite (RM) simple sequence repeats (SSR’S) is a pre-requisite for genotypic screening to identify the loci associated with trait of interest among mapping population. In the present study, eight popularly used rice maintainer lines viz., APMS-6B (Improved for Bacterial leaf blight. [1], IR58025B, IR68897B, IR79156B, DRR-6B, DRR-9B, BF-16B and BF2096B were used to study stigma exsertion trait as a single, double and total stigma exsertion. A total of 630RM markers were used to study parental polymorphism among eight maintainer lines and also to map their association with stigma exsertion trait. Among 630, 253 RM markers showed polymorphism with 635 alleles among the eight maintainers which were distributed across twelve chromosomes of rice. The overall parental survey revealed 40.18 per cent of polymorphism among the maintainer lines with a maximum and minimum frequency of 5 and 2 alleles, respectively. The genetic similarity coefficient for the most number of pairs ranged between of 0.2-0.9 with the average value of 0.60 for all possible combinations, indicating moderate genetic diversity among the chosen genotypes. The genotypes grouped according to their place of origin and represents genetic closeness between them. The identified RM polymorphic markers could be used to construct the linkage map and subsequently, to identify the stigma exsertion related QTLs from mapping population developed from different combinations of the rice maintainer lines.

Microsatellites based parental polymorphism survey for moisture stress in Rice (Oryza sativa L.) between the parental genotypes HUR-1309 and CR Dhan 801

The present investigation was conducted during Rabi 2018-19. The parental polymorphism research was done at the Molecular Drought Breeding Laboratory (Department of Genetics and Plant Breeding), Institute of Agricultural Sciences, BHU, and the Crop Improvement Division, ICAR-NRRI, Cuttack. The purpose of this research was to identify the parental polymorphism markers, analyze their chromosomal distribution, and determine the repetitive motifs. The polymorphic markers will be helpful to introgress the drought tolerant yield QTLs (qDTY) from the donor CR Dhan 801 into the background of HUR-1309, a popular aromatic short duration variety through marker-assisted backcross breeding program. Polymerase chain reaction (PCR), agarose gel electrophoresis, and genomic DNA separation were carried out according to standard procedures. A total of 510 microsatellites were used for evaluation of the parental polymorphism and 90 markers showed polymorphism among the parents and the rest 420 markers are monomorphic for the studied QTLs (qDTY1.1, qDTY2.1, and qDTY3.1). The parental polymorphism in the present study ranged between 11.43% to 30.61%. Considering the 12 chromosomes of rice, the average polymorphism was observed 17.65%. Out of 90 polymorphic markers, 59 were dinucleotide repeats, 24 were trinucleotide repeats and 4 were tetra nucleotide repeats. The dinucleotides were highly present on chromosomes 1 (12), 2 (9), and 3 (9) and repeated 12, 9, and 9 times respectively. The markers which are found to be polymorphic can be used in the marker-assisted backcross breeding (MABB) program for grain yield under moisture stress.

Fine mapping of <i>Rf5</i> region for a sorghum fertility restorer gene and microsynteny analysis across grass species

Cytoplasmic male sterility (CMS) is widely used to control pollination in the production of commercial F1 hybrid seed in sorghum. So far, 6 major fertility restorer genes, Rf1 to Rf6, have been reported in sorghum. Here, we fine-mapped the Rf5 locus on sorghum chromosome 5 using descendant populations of a ‘Nakei MS-3A’ × ‘JN43’ cross. The Rf5 locus was narrowed to a 140-kb region in BTx623 genome (161-kb in JN43) with 16 predicted genes, including 6 homologous to the rice fertility restorer Rf1 (PPR.1 to PPR.6). These 6 homologs have tandem pentatricopeptide repeat (PPR) motifs. Many Rf genes encode PPR proteins, which bind RNA transcripts and modulate gene expression at the RNA level. No PPR genes were detected at the Rf5 locus on the corresponding homologous chromosome of rice, foxtail millet, or maize, so this gene cluster may have originated by chromosome translocation and duplication after the divergence of sorghum from these species. Comparison of the sequences of these genes between fertile and CMS lines identified PPR.4 as the most plausible candidate gene for Rf5.

The induced mutant allele flo4-303 confers floury characteristics on the japonica rice cultivar 'Hoshinoko'.

Rice flour is useful as a substitute for wheat flour, however, to obtain fine flour, millers need special milling facilities, which increase the cost of milling. To reduce the milling cost, we developed a floury mutant line by irradiating gamma-rays to dry seeds of the japonica cultivar 'Hoshinoyume'. The line was registered as a new cultivar, 'Hoshinoko'. Genetical analysis of the floury trait was conducted using an F2 population derived from a cross between 'Hoshinoko' and 'Corbetti' (a japonica rice cultivar with normal endosperm), which indicated the involvement of a single recessive gene located near the RM163 marker on the long arm of rice chromosome 5, flanking flo4 identified by Kang et al. (2005). Sequence analysis of flo4 showed a two-bp (CA) insertion in the eighth exon of in 'Hoshinoko' compared to that of 'Hoshinoyume', which led to a frameshift mutation. The CAPS-based genotype of flo4 gene completely correlated to the phenotype of endosperm in two populations. This CAPS marker could be helpful for rice breeders to develop new cultivars harboring floury endosperm of the flo4-303 gene.

Study of Parental Polymorphism and Allelic Variation for Grain Quality and Yield Traits in Rice (Oryza sativa L.) Using SSR Markers

Aim: Identification of polymorphic markers is prerequisite for conducting any QTL mapping experiment because if the parents are polymorphic for the traits of interest, then further selection of plants in the progenies becomes easy. Hence, the objective of the present study was to identify polymorphic markers for grain quality and yield traits among the parental lines Improved Samba Mahsuri and Badshabhog.
 Place and Duration of Study: It was carried out at Molecular Breeding Lab, Department of Genetics and Plant Breeding, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi - 221 005, India, during 2019.
 Methodology: Two parents Improved Samba Mahsuri and Badshabhog were used for the present study. The DNA extraction was done as per the CTAB method suggested by Murray and Thompson. Standard PCR protocol was followed.
 Results: For parental polymorphism survey, a total of 576 randomly selected SSR markers including 26 gene specific markers related to aroma, cooking and eating quality, grain dimension and yield related traits distributed across the 12 chromosomes of rice were used. Overall, 96 markers including 4 gene specific markers were found to be polymorphic between the two genotypes indicating a total polymorphism percentage of 16.67%. The highest polymorphism percentage was recorded on chromosome 6 (26.67%) followed by chromosome 4 (21.43%) and the lowest polymorphism percentage was observed on chromosome 10 (8.93%). The gene specific markers nksbad2, ARO7, BADEX7_5 and SSI were found to be polymorphic.
 Conclusion: Based on the present study it may be concluded that the polymorphic markers identified will further be utilized in genotyping of F2:3 population, linkage analysis and mapping QTL’s for grain quality and yield traits.

A Rice Ancestral Genetic Resource Conferring Ideal Plant Shapes for Vegetative Growth and Weed Suppression.

To maximize crop growth, crops need to capture sunlight efficiently. This property is primarily influenced by the shape of the crops such as the angle, area, and arrangement of leaves. We constructed a rice (Oryza sativa L.) inbred line that displayed an ideal transition of plant shapes in terms of sunlight receiving efficiency. During vegetative growth, this line exhibited tiller spreading with increased tiller number, which formed a parabolic antenna-like structure. The architecture probably improved light reception efficiency of individuals compared with the recurrent parent. The line achieved not only acceleration of the vegetative growth, but also significant suppression of weed growth under the canopy. The increased light reception efficiency of the line has consequently reduced the amount of incident light to the ground and supplied significant competitiveness against weeds. The spread tillers became erect from the entry of the reproductive growth phase, adaptively sustaining light reception efficiency in thicker stands. The line carries a small chromosomal segment from Oryza rufipogon Griff., a putative progenitor of Asian cultivated rice. The introduced chromosome segment had little effect on grain yield and quality. Our results shed light on potentials hidden in the wild rice chromosome segment to achieve the valuable traits.

Identification and verification of grain shape QTLs by SNP array in rice.

Grain shape strongly influences the economic value and grain yield of rice. Thus, identifying quantitative trait loci (QTLs) for grain shape has been a longstanding goal in rice genetic research and breeding programs. Single nucleotide polymorphism (SNP) markers are ubiquitous in the rice genome and are more abundant and evenly distributed on the 12 rice chromosomes than traditional markers. An F2 population was genotyped using the RICE6K SNP array to elucidate the mechanisms governing grain shape. Thirty-five QTLs for grain shape were detected on 11 of 12 chromosomes over 2 years. The major QTL cluster qGS7 was detected in both years and displayed strong genetic effects on grain length and width, showing consistency with GL7/GW7. Some minor QTLs were also detected, and the effects of four QTLs on seed size were then validated using BC1F6 populations with residual heterozygous lines in each QTL region. Our findings provide insights into the molecular basis of grain shape as well as additional resources and approaches for producing hybrid high-yield rice varieties.

Genome-wide SNP discovery, linkage mapping, and analysis of QTL for morpho-physiological traits in rice during vegetative stage under drought stress.

The online version contains supplementary material available at 10.1007/s12298-021-01095-y.

Breeding for Low Temperature Germinability in Temperate Japonica Rice Varieties: Analysis of Candidate Genes in Associated QTLs

In temperate areas, rice deals with low temperatures that can affect plant growth and crop yield. Rapid germination is required for adequate plant establishment in the field, therefore obtaining cultivars that maintain this phenotype under suboptimal temperature conditions is a challenge for rice breeders. Our study aimed to investigate temperature-induced expression changes in genes underlying quantitative trait loci (QTLs) associated to this trait (low temperature germinability, LTG) that were detected in a previous genome wide association study (GWAS). In the context of a breeding program for japonica rice cultivars adapted to cultivation in Spain, we obtained two biparental families of lines derived from hybridization with two cold tolerant Italian cultivars, and we have studied the effect on the LTG phenotype of introgressing these QTLs. A wide region in chromosome 3 was related to significant increases in seedling growth rate at 15 °C, although the extent of the effect depended on the analyzed family. In parallel, we studied the pattern of expression during germination at different temperatures of 10 genes located in the LTG-associated QTLs, in five japonica rice cultivars and in a biparental family of recombinant inbred lines (RILs). Cold induced changes in the expression of the 10 analyzed genes, with significant differences among genotypes. Variation in LTG phenotype was consistently associated with changes in the pattern of expression of five genes from the tagged regions in rice chromosome 3, which encoded for enzymes implicated in phytohormone metabolism (OsFBK12, Os3Bglu6), oxidative stress (SPL35, OsSRO1c) and Mn homeostasis maintenance (OsMTP8.1). Differential expression induced by cold in two regulatory genes (Os02g0824000 and Os06g06400) also contributed to explain low temperature tolerance during rice germination. In conclusion, introgression in defective cultivars of favorable alleles for these genes would contribute to the genetic improvement of LTG in japonica rice varieties.

Pinpointing genomic regions associated with root system architecture in rice through an integrative meta-analysis approach.

Applying an integrated meta-analysis approach led to identification of meta-QTLs/ candidate genes associated with rice root system architecture, which can be used in MQTL-assisted breeding/ genetic engineering of root traits. Root system architecture (RSA) is an important factor for facilitating water and nutrient uptake from deep soils and adaptation to drought stress conditions. In the present research, an integrated meta-analysis approach was employed to find candidate genes and genomic regions involved in rice RSA traits. A whole-genome meta-analysis was performed for 425 initial QTLs reported in 34 independent experiments controlling RSA traits under control and drought stress conditions in the previous twenty years. Sixty-four consensus meta-QTLs (MQTLs) were detected, unevenly distributed on twelve rice chromosomes. The confidence interval (CI) of the identified MQTLs was obtained as 0.11-14.23cM with an average of 3.79cM, which was 3.88 times narrower than the mean CI of the original QTLs. Interestingly, 52 MQTLs were co-located with SNP peak positions reported in rice genome-wide association studies (GWAS) for root morphological traits. The genes located in these RSA-related MQTLs were detected and explored to find the drought-responsive genes in the rice root based on the RNA-seq and microarray data. Multiple RSA and drought tolerance-associated genes were found in the MQTLs including the genes involved in auxin biosynthesis or signaling (e.g. YUCCA, WOX, AUX/IAA, ARF), root angle (DRO1-related genes), lateral root development (e.g. DSR, WRKY), root diameter (e.g. OsNAC5), plant cell wall (e.g. EXPA), and lignification (e.g. C4H, PAL, PRX and CAD). The genes located within both the SNP peak positions and the QTL-overview peaks for RSA are suggested as novel candidate genes for further functional analysis. The promising candidate genes and MQTLs can be used as basis for genetic engineering and MQTL-assisted breeding of root phenotypes to improve yield potential, stability and performance in a water-stressed environment.

Development and regeneration of wheat-rice hybrid zygotes produced by in vitro fertilization system.

Summary Hybridization plays a decisive role in the evolution and diversification of angiosperms. However, the mechanisms of wide hybridization remain open because pre‐ and post‐fertilization barriers limit the production and development of inter‐subfamily/intergeneric zygotes, respectively.We examined hybridization between wheat and rice using an in vitro fertilization (IVF) system to bypass these barriers. Several gamete combinations of allopolyploid wheat–rice hybrid zygotes were successfully produced, and the developmental profiles of hybrid zygotes were analyzed.Hybrid zygotes derived from one rice egg cell and one wheat sperm cell ceased at the multicellular embryo‐like structure stage. This developmental barrier was overcome by adding one wheat egg cell to the wheat–rice hybrid zygote. In the reciprocal combination, one wheat egg and one rice sperm cell, the resulting hybrid zygotes failed to divide. However, doubling the dosage of rice sperm cell allowed the hybrid zygotes to develop into plantlets. Rice chromosomes appeared to be progressively eliminated during the early developmental stage of these hybrid embryos, and c. 20% of regenerated plants showed abnormal morphology.These results suggest that hybrid breakdown can be overcome through optimization of gamete combinations, and the present hybrid will provide a new horizon for utilization of inter‐subfamily genetic resources.

Substitution Mapping of Two Closely Linked QTLs on Chromosome 8 Controlling Grain Chalkiness in Rice

Rice varieties are required to have high yield and good grain quality. Grain chalkiness and grain shape are two important traits of rice grain quality. Low chalkiness slender grains are preferred by most rice consumers. Here, we dissected two closely linked quantitative trait loci (QTLs) controlling grain chalkiness and grain shape on rice chromosome 8 by substitution mapping. Two closely linked QTLs controlling grain chalkiness and grain shape were identified using single-segment substitution lines (SSSLs). The two QTLs were then dissected on rice chromosome 8 by secondary substitution mapping. qPGC8.1 was located in an interval of 1382.6 kb and qPGC8.2 was mapped in a 2057.1 kb region. The maximum distance of the two QTLs was 4.37 Mb and the space distance of two QTL intervals was 0.72 Mb. qPGC8.1 controlled grain chalkiness and grain width. qPGC8.2 was responsible for grain chalkiness, grain length and width. The additive effects of qPGC8.1 and qPGC8.2 on grain chalkiness were not affected by higher temperature. Two closely linked QTLs qPGC8.1 and qPGC8.2 were dissected on rice chromosome 8. They controlled the phenotypes of grain chalkiness and grain shape. The two QTLs were insensitive to higher temperature.

Karyotype diversity in cultivated and wild Indian rice through EMA-based chromosome analysis

India is known for its diverse cultivated and wild rice germplasm. In today's crop improvement programmes, wild relatives are much-needed genetic repository of valuable traits. Analysis of genetic diversity at the chromosomal level is one cost-effective tool to unlock foundational information related to genetics and plant breeding. Presently, enzymatic maceration and air-drying method (EMA) has been applied for the first time in six cultivated and nine wild Indian rice (diploid and tetraploid). EMA method following Giemsa staining has yielded large numbers of cytoplasm free metaphase plates with distinct chromosome morphology. Detailed analysis has revealed karyotype diversities in terms of total chromatin length (TCL), chromosome morphology and location of sat chromosomes within and between the studied species. Most of the cultivated rice has gained additional amount in TCL during the period of domestication in comparison to their progenitor Oryza nivara. Morphological clarity of the small chromosomes of rice was much required and has helped to identify individual chromosomes in the diverse karyotypes. Diversity in landmark SAT chromosomes is another important observation, not reported previously in Indian rice. Present study has shown that in most of the O. sativa members, the 10th pair contains SAT except one where 6th pair is satellited. On the other hand, diversity of SAT in diploid and tetraplod wild species has been recorded on 5th, 7th and 8th chromosome pairs and on 9th, 12th, 22nd and 23rd chromosome pairs, respectively. Karyomorphometric indices has helped to construct dendrogram to elucidate intraspecies and interspecies relationships. Untapped genetic diversity recorded in Indian rice through chromosomal analysis will be useful to the breeders and genome researchers.

Recent Advances in Rice Varietal Development for Durable Resistance to Biotic and Abiotic Stresses through Marker-Assisted Gene Pyramiding

Abiotic and biotic stresses adversely affect rice growth, development and grain yield. Traditional rice breeding techniques are insufficient in modern agriculture to meet the growing population’s food needs on a long-term basis. The development of DNA markers closely linked to target genes or QTLs on rice chromosomes, and advanced molecular techniques, such as marker-assisted selection (MAS), have encouraged the evolution of contemporary techniques in rice genetics and breeding, such as gene pyramiding. Gene pyramiding refers to the act of combining two or more genes from multiple parents into a single genotype, which allows the overexpression of more than one gene for broad-spectrum abiotic and biotic stress resistance. Marker-assisted pedigree, backcrossing and pseudo-backcrossing methods can increase the conventional breeding speed by reducing the number of breeding generations in order to enhance the pyramiding process. Pyramiding is affected by several factors: the number of transferred genes; the range within gene and flanking markers; the number of chosen populations in every breeding generation; the features of genes and germplasms; and the potentiality of breeders to identify the target genes. Modern breeding methods, such as the marker-assisted backcrossing approach, have made gene pyramiding more precise and reliable for the development of stress-tolerant rice varieties in the coming decades. This review presents up-to-date knowledge on gene pyramiding schemes, marker-assisted gene pyramiding techniques, the efficiency of marker-assisted gene pyramiding and the advantages and limitations of gene pyramiding methods. This review also reports on the potential application of marker-assisted selection breeding to develop stress-tolerant rice varieties that stabilize abiotic and biotic stresses. This review will help rice breeders to improve yields by increasing rice productivity under abiotic and biotic stress conditions.