Stay Green Research Articles

UAV-Based Phenotyping: A Non-Destructive Approach to Studying Wheat Growth Patterns for Crop Improvement and Breeding Programs

Rising food demands require new techniques to achieve higher genetic gains for crop production, especially in regions where climate can negatively affect agriculture. Wheat is a staple crop that often encounters this challenge, and ideotype breeding with optimized canopy traits for grain yield, such as determinate tillering, synchronized flowering, and stay-green (SG), can potentially improve yield under terminal drought conditions. Among these traits, SG has emerged as a key factor for improving grain quality and yield by prolonging photosynthetic activity during reproductive stages. This study aims to highlight the importance of growth dynamics in a wheat mapping population by using multispectral images obtained from uncrewed aerial vehicles as a high-throughput phenotyping technique to assess the effectiveness of using such images for determining correlations between vegetation indices and grain yield, particularly regarding the SG trait. Results show that the determinate group exhibited a positive correlation between NDVI and grain yield, indicating the effectiveness of these traits in yield improvement. In contrast, the indeterminate group, characterized by excessive vegetative growth, showed no significant NDVI–grain yield relationship, suggesting that NDVI values in this group were influenced by sterile tillers rather than contributing to yield. These findings provide valuable insights for crop breeders by offering a non-destructive approach to enhancing genetic gains through the improved selection of resilient wheat genotypes.

CfSGR1 and CfSGR2 from Cryptomeria fortunei exhibit contrasting responses to hormones and abiotic stress in transgenic Arabidopsis

Stay-green (SGR) genes are pivotal regulatory genes in the context of plant chlorophyll metabolism, but few studies on SGR homologues in Cryptomeria fortunei have been previously reported. We cloned two CfSGR genes and overexpressed them in Arabidopsis to explore their functions. Full-length CfSGR1 and CfSGR2 are 1265 and 1197 bp, encompassing open reading frames (ORFs) encoding 274 and 276 amino acids, respectively. SGRs exhibited high conservation in higher plants, and phylogenetic analysis indicated that SGRs from monocots and gymnosperms cluster in a clade. The proteins localized to chloroplasts and showed no transcriptional activity in yeast cells. The CfSGR gene expressions were induced by abiotic stresses and hormones. Under conditions of darkness, abscisic acid (ABA), salt, drought, or freezing stress, CfSGR2-transgenic Arabidopsis exhibited a delay in leaf yellowing compared to the WT, which was attributed to increased chlorophyll content and enhanced photosynthetic capacity. These transgenic plants exhibited improved resistance to stress via upregulated expression of resistance-related genes, increased antioxidant enzyme activities, and reduced malondialdehyde content and electrolyte leakage rate. In contrast, CfSGR1-transgenic plants may accelerate leaf yellowing and exhibit reduced stress resistance. Our findings highlight potential divergence in the functions of CfSGR genes concerning plant growth and development and responses to abiotic stresses or hormones, providing a scientific foundation for future breeding of stress-resistant C. fortunei cultivars.

Activity examination of plant Mg-dechelatase and its bacterial homolog in plants and in vitro

Chlorophyll a serves as a photosynthetic pigment in plants. Its degradation is initiated by the extraction of the central Mg by the Mg-dechelatase enzyme, which is encoded by Stay-Green (SGR). Plant SGR is believed to be derived from bacterial SGR homolog obtained through horizontal gene transfer into photosynthetic eukaryotes. However, it is not known how the bacterial SGR homolog was modified to function in plants. To assess its adaptation mechanism in plants, a bacterial SGR homolog derived from the Anaerolineae bacterium SM23_63 was introduced into plants. It was found that the bacterial SGR homolog metabolized chlorophyll in plants. However, its chlorophyll catabolic activity was lower than that of plant SGR. Recombinant proteins of the bacterial SGR homolog exhibited higher activity than those of the plant SGR. The reduced chlorophyll catabolic activity of bacterial SGR homologs in plants may be associated with low hydrophobicity of the entrance to the catalytic site compared to that of plant SGR. This hinders the enzyme access to chlorophyll, which is localized in hydrophobic environments. This study offers insights into the molecular changes underlying the optimization of enzyme function.

RsOBP2a, a member of OBF BINDING PROTEIN transcription factors, inhibits two chlorophyll degradation genes in green radish

The green radish (Raphanus sativus L.) contains abundant chlorophyll (Chl). DOF-type transcription factor OBF BINDING PROTEIN (OBP) plays crucial functions in plant growth, development, maturation and responses to various abiotic stresses. However, the metabolism by which OBP transcription factors regulate light-induced Chl metabolism in green radish is not well understood. In this study, six OBP genes were identified from the radish genome, distributed unevenly across five chromosomes. Among these genes, RsOBP2a showed significantly higher expression in the green flesh compared to the white flesh of green radish. Analysis of promoter elements suggested that RsOBPs might be involved in stress responses, particularly in light-related processes. Overexpression of RsOBP2a led to increase Chl levels in cotyledons and adventitious roots of radish, while silencing RsOBP2a expression through TYMV-induced gene silencing accelerated leaf senescence. Further investigations revealed that RsOBP2a was localized in the nucleus and served as a transcriptional repressor. RsOBP2a was induced by light and directly suppressed the expression of STAYGREEN (SGR) and RED CHLOROPHYLL CATABOLITE REDUCTASE (RCCR), thereby delaying senescence in radish. Overall, a novel regulatory model involving RsOBP2a, RsSGR, and RsRCCR was proposed to govern Chl metabolism in response to light, offering insights for the enhancement of green radish germplasm.

Genome-Wide Identification and Characterization of Tea SGR Family Members Reveal Their Potential Roles in Chlorophyll Degradation and Stress Tolerance

Photosynthesis plays vital roles in plant growth and development. Stay-green (SGR) proteins are responsible for chlorophyll degradation and photosynthetic metabolism. To identify SGR family members and determine their potential functions in tea plants, we identified and cloned three SGR genes. Phylogenetic analysis revealed that the tea SGR homologs were classified into the SGR subfamily (named CsSGR1 and CsSGR2) and the SGRL subfamily (named CsSGRL). Cis-element analysis indicated that the promoters of CsSGR1, CsSGR2 and CsSGRL contained light-, phytohormone- and stress-related elements. Subcellular localization confirmed that CsSGR1 was localized in the chloroplast, while CsSGR2 and CsSGRL were localized in the chloroplast, membrane and nucleus. The RT-qPCR results showed that the three genes in the matures of albino tea cultivars were expressed higher than in the green tea cultivar. However, only CsSGR2-overexpressing tobacco leaves exhibited a yellowish phenotype and significantly lower Fv/Fm values. CsSGR1 and CsSGR2 exhibited similar expression patterns in different tissues after infection with the pathogen Colletotrichum camelliae, which was opposite to the pattern observed for CsSGRL. In addition, CsSGR1 was significantly induced in response to cold stress, SA, JA and ABA in C. camelliae. These findings identified valuable candidate genes for elucidating the mechanism of leaf albinism, stress response and phytohormone signaling in tea plants.

Transcriptome Sequencing Provides Insights into High-Temperature-Induced Leaf Senescence in Herbaceous Peony

Global warming causes frequent high temperatures in summer; which negatively impacts herbaceous peonies (Paeonia lactiflora Pall.) by accelerating leaf senescence and reducing biomass accumulation, leading to reduced flower quality in the subsequent year. Our findings revealed that as heat stress progressed, the high-temperature-sensitive cultivar ‘Meigui Zi’ (MGZ) exhibited a higher rate of chlorophyll content reduction and more pronounced premature aging symptoms than the high-temperature-tolerant cultivar ‘Chi Fen’ (CF). To investigate gene expression differences between CF and MGZ under high-temperature stress, we combined PacBio Iso-Seq sequencing (Iso-Seq) with next-generation sequencing (RNA-seq). Iso-seq yielded 352,891 full-length transcripts ranging from 61 bp to 49,022 bp in length. RNA-seq generated 257,562 transcripts across all samples. Further analysis revealed that differentially expressed genes (DEGs) between CF and MGZ were primarily enriched in “Photosynthesis”, with most photosynthesis-related DEGs highly expressed in CF. This indicates that CF has higher stability in its photosystem compared with MGZ, which is crucial for mitigating leaf senescence caused by high temperatures. Additionally, the highly expressed chlorophyll degradation genes stay-green (SGR) and stay-green-like (SGRL) in MGZ may be involved in chlorophyll content reduction induced by high temperature. This study preliminarily revealed the molecular mechanism of high-temperature-induced leaf senescence of in herbaceous peony and provided candidate genes for further studies of the regulation mechanism of high -temperature-induced leaf senescence.

Using UAV-Based Temporal Spectral Indices to Dissect Changes in the Stay-Green Trait in Wheat.

Stay-green (SG) in wheat is a beneficial trait that increases yield and stress tolerance. However, conventional phenotyping techniques limited the understanding of its genetic basis. Spectral indices (SIs) as non-destructive tools to evaluate crop temporal senescence provide an alternative strategy. Here, we applied SIs to monitor the senescence dynamics of 565 diverse wheat accessions from anthesis to maturation stages over 2 field seasons. Four SIs (normalized difference vegetation index, green normalized difference vegetation index, normalized difference red edge index, and optimized soil-adjusted vegetation index) were normalized to develop relative stay-green scores (RSGS) as the SG indicators. An RSGS-based genome-wide association study identified 47 high-confidence quantitative trait loci (QTL) harboring 3,079 single-nucleotide polymorphisms associated with SG and 1,085 corresponding candidate genes. Among them, 15 QTL overlapped or were adjacent to known SG-related QTL/genes, while the remaining QTL were novel. Notably, a set of favorable haplotypes of SG-related candidate genes such as TraesCS2A03G1081100, TracesCS6B03G0356400, and TracesCS2B03G1299500 are increasing following the Green Revolution, further validating the feasibility of the pipeline. This study provided a valuable reference for further quantitative SG and genetic research in diverse wheat panels.

Insights into the physiological, molecular, and genetic regulators of albinism in Camellia sinensis leaves.

Introduction: Yanling Yinbiancha, a cultivar of Camellia sinensis (L.) O. Kuntze, is an evergreen woody perennial with characteristic albino leaves. A mutant variant with green leaves on branches has been recently identified. The molecular mechanisms underlying this color variation remain unknown. Methods: We aimed to utilize omics tools to decipher the molecular basis for this color variation, with the ultimate goal of enhancing existing germplasm and utilizing it in future breeding programs. Results and discussion: Albinotic leaves exhibited significant chloroplast degeneration and reduced carotenoid accumulation. Transcriptomic and metabolomic analysis of the two variants revealed 1,412 differentially expressed genes and 127 differentially accumulated metabolites (DAMs). Enrichment analysis for DEGs suggested significant enrichment of pathways involved in the biosynthesis of anthocyanins, porphyrin, chlorophyll, and carotenoids. To further narrow down the causal variation for albinotic leaves, we performed a conjoint analysis of metabolome and transcriptome and identified putative candidate genes responsible for albinism in C. sinensis leaves. 12, 7, and 28 DEGs were significantly associated with photosynthesis, porphyrin/chlorophyll metabolism, and flavonoid metabolism, respectively. Chlorophyllase 2, Chlorophyll a-Binding Protein 4A, Chlorophyll a-Binding Protein 24, Stay Green Regulator, Photosystem II Cytochrome b559 subunit beta along with transcription factors AP2, bZIP, MYB, and WRKY were identified as a potential regulator of albinism in Yanling Yinbiancha. Moreover, we identified Anthocyanidin reductase and Arabidopsis Response Regulator 1 as DEGs influencing flavonoid accumulation in albino leaves. Identification of genes related to albinism in C. sinensis may facilitate genetic modification or development of molecular markers, potentially enhancing cultivation efficiency and expanding the germplasm for utilization in breeding programs.

A hypervariable intron of the STAYGREEN locus provides excellent discrimination among Pisum fulvum accessions and reveals evidence for a relatively recent hybridization event with Pisum sativum.

An analysis of 82 non-synonymous Pisum fulvum accessions for sequence variation in a fragment of the STAYGREEN (SGR) locus revealed 57 alleles, most of which differed in indel structure. Eight additional P. fulvum accessions, each supposedly synonymous with a different accession of the initial group, were also analyzed. In every case the paired synonymous accessions possessed the same SGR sequence but varied slightly for a 6-trait morphological phenotype, indicating that SGR sequence is a much more reliable indicator of accession identity than is a morphological characterization. SGR sequence analysis confirmed our previous finding that P. fulvum accessions separate into two allele groups. This division was not supported by results of previous studies that were based on sequences distributed across the entire genome, suggesting that the division may have been produced by selection at a nearby locus and that the SGR phylogeny may not be good indicator of overall relationships within the species. One P. fulvum accession, PI 595941 (=JI1796), displayed an SGR sequence outside the variation typical of the species. Instead, its allele resembled alleles limited to a set of Pisum sativum landraces from the Middle East, suggesting hybridization between ancestors of PI 595941 and some primitive form of domesticated P. sativum. With one exception from the extreme northwest corner of Israel, P. fulvum accessions collected north of latitude 35.5° N were fixed for alleles from group A. These northern accessions also displayed greatly reduced SGR sequence diversity compared to group A accessions collected from other regions, suggesting that the northern populations may represent recent extensions of the range of the species. Group B accessions were distributed from Lake Tiberias south and were generally sympatric with the southern group A accessions. Although group B accessions occupied a smaller area than group A, the SGR sequence diversity in this group (28 alleles in 33 accessions) exceeded that for group A.

Introgression of Stay Green Quantitative Trait Locus (QTLS) into Elite Sorghum Variety by MABC

Sorghum is a major staple food crop for the people in semi-arid areas of Asia. Post-flowering drought is a global constraint of sorghum production. The study aimed to improve stay-green characteristics of GS-23 sorghum variety by transferring stg3A and stg3B QTL`s respectively from donor genotypes K260 and K359w of ICRISAT by using marker assisted backcrossing. The experimental material comprised of six basic generations P1, P2, F1, F2, BC1 and BC2 developed from crossing GS-23 × K260 and GS-23 × K359 were genotyped using a set of 133 SNPs and 79 SSR markers. Whereas, 53 polymorphic SNPs among parents and backcross F1s for stay green trait at maturity were used to track introgression of stay green trait. Similarly,10 SSR markers were found to be polymorphic were used to track introgression of stay green trait i.e., stay green trait QTL`s stg3A and stg3B from donor parent K260 and K359w respectively in GS-23 background across backcross population and to identify plants that were homozygous for the desired allele. In which 02 SNPs SnpSB0039 and SnpSB0093 were identified as polymorphic for both K260 and K359w. Whereas 17 SNPs identified polymorphic for K260 and 34 SNPs for K359w. These SNPs were validated in both F1 and BC1F1 populations of both the crosses. Similarly, out of 10 SSR markers utilized, 02 SSRs were identified polymorphic to K260 and 02 SSRs were identified for K359w. Whereas 06 SSRs were polymorphic to both the parents K260 and K359w. Prominent 02 SSRs viz., Xtxp 141_Fam and Xgap84_Vic found to be more reliable and polymorphic to both the parents. These SSRs were validated in both F1 and BC1F1 populations of both the crosses. The genotypic analysis revealed the presence of favorable alleles in homozygous conditions at markers loci associated with stg3A and stg3B QTL`s in BC populations, suggesting successful introgression of stay green QTLs from the donor parents to the recurrent parent. Therefore, our study demonstrated the utility of marker-assisted backcrossing for drought tolerance improvement of locally adapted sorghum variety.

Receptor for Activated C Kinase1B (RACK1B) Delays Salinity-Induced Senescence in Rice Leaves by Regulating Chlorophyll Degradation.

The widely conserved Receptor for Activated C Kinase1 (RACK1) protein is a WD-40 type scaffold protein that regulates diverse environmental stress signal transduction pathways. Arabidopsis RACK1A has been reported to interact with various proteins in salt stress and Light-Harvesting Complex (LHC) pathways. However, the mechanism of how RACK1 contributes to the photosystem and chlorophyll metabolism in stress conditions remains elusive. In this study, using T-DNA-mediated activation tagging transgenic rice (Oryza sativa L.) lines, we show that leaves from rice RACK1B gene (OsRACK1B) gain-of-function (RACK1B-OX) plants exhibit the stay-green phenotype under salinity stress. In contrast, leaves from down-regulated OsRACK1B (RACK1B-UX) plants display an accelerated yellowing. qRT-PCR analysis revealed that several genes which encode chlorophyll catabolic enzymes (CCEs) are differentially expressed in both RACK1B-OX and RACK1B-UX rice plants. In addition to CCEs, stay-green (SGR) is a key component that forms the SGR-CCE complex in senescing chloroplasts, and which causes LHCII complex instability. Transcript and protein profiling revealed a significant upregulation of OsSGR in RACK1B-UX plants compared to that in RACK1B-OX rice plants during salt treatment. The results imply that senescence-associated transcription factors (TFs) are altered following altered OsRACK1B expression, indicating a transcriptional reprogramming by OsRACK1B and a novel regulatory mechanism involving the OsRACK1B-OsSGR-TFs complex. Our findings suggest that the ectopic expression of OsRACK1B negatively regulates chlorophyll degradation, leads to a steady level of LHC-II isoform Lhcb1, an essential prerequisite for the state transition of photosynthesis for adaptation, and delays salinity-induced senescence. Taken together, these results provide important insights into the molecular mechanisms of salinity-induced senescence, which can be useful in circumventing the effect of salt on photosynthesis and in reducing the yield penalty of important cereal crops, such as rice, in global climate change conditions.

Water stress effects on stay green and chlorophyll fluorescence with focus on yield characteristics of diverse bread wheats.

The study provided an insight toward better understanding of stay-green mechanisms for drought tolerance improvement and identified that synthetic-derived wheats proved as a promising germplasm for improved tolerance against water stress. Stay-green (SG) trait is considered to be related with the ability of wheat plants to maintain photosynthesis and CO2 assimilation. The present study explored the interaction of water stress with SG expression through physio-biochemical, agronomic and phenotypic responses among diverse wheat germplasm comprising of 200 synthetic hexaploids, 12 synthetic derivatives, 97 landraces and 16 conventional bread wheat varieties, for 2years. The study established that variation of SG trait existed in the studied wheat germplasm and there was positive association between SG trait and tolerance to water stress. The relationship of SG trait with chlorophyll content (r = 0.97), ETR (r = 0.28), GNS (r = 0.44), BMP (r = 0.34) and GYP (r = 0.44) was particularly promising under water stress environment. Regarding chlorophyll fluorescence, the positive correlation of фPSII (r = 0.21), qP (r = 0.27) and ETR (r = 0.44) with grain yield per plant was noted. The improved ΦPSII and Fv/Fm of PSII photochemistry resulted in the high photosynthesis activity in SG wheat genotypes. Regarding relative water content and photochemical quenching coefficient, synthetic-derived wheats were better by maintaining 20.9, 9.8 and 16.1% more RWC and exhibiting 30.2, 13.5 and 17.9% more qP when compared with landraces, varieties and synthetic hexaploids, respectively, under water stress environment. Synthetic derived wheats also exhibited relatively more SG character with good yield and were more tolerant to water stress in terms of grain yield, grain weight per plant, better photosynthetic performance through chlorophyll fluorescence measurement, high leaf chlorophyll and proline content, and hence, may be used as novel sources for breeding drought tolerant materials. The study will further facilitate research on wheat leaf senescence and will add to better understanding of SG mechanisms for drought tolerance improvement.

Genome-Wide Identification of Kiwifruit SGR Family Members and Functional Characterization of SGR2 Protein for Chlorophyll Degradation.

The STAY-GREEN (SGR) proteins play an important role in chlorophyll (Chl) degradation and are closely related to plant photosynthesis. However, the availability of inadequate studies on SGR motivated us to conduct a comprehensive study on the identification and functional dissection of SGR superfamily members in kiwifruit. Here, we identified five SGR genes for each of the kiwifruit species [Actinidia chinensis (Ac) and Actinidia eriantha (Ae)]. The phylogenetic analysis showed that the kiwifruit SGR superfamily members were divided into two subfamilies the SGR subfamily and the SGRL subfamily. The results of transcriptome data and RT-qPCR showed that the expression of the kiwifruit SGRs was closely related to light and plant developmental stages (regulated by plant growth regulators), which were further supported by the presence of light and the plant hormone-responsive cis-regulatory element in the promoter region. The subcellular localization analysis of the AcSGR2 protein confirmed its localization in the chloroplast. The Fv/Fm, SPAD value, and Chl contents were decreased in overexpressed AcSGR2, but varied in different cultivars of A. chinensis. The sequence analysis showed significant differences within AcSGR2 proteins. Our findings provide valuable insights into the characteristics and evolutionary patterns of SGR genes in kiwifruit, and shall assist kiwifruit breeders to enhance cultivar development.

Characterisation and evaluation of wheat genetic resources for heat stress tolerance using stay-green traits

Context Post-anthesis heat stress is a major concern for wheat. Stay-green (SG) can serve as a crucial marker for plant adaptation to it. Though genetic resources provide an invaluable gene pool for crop breeding, collections are still uncharacterised and their potential is yet to be explored. Aims This study was planned to characterise and evaluate wheat genetic resources, including wild germplasm, for heat stress tolerance using SG traits as selection criteria. Methods Experiment was conducted with thirty wheat genotypes under late and very late sown environments for 2 years. Genotypes were assessed for SG traits like Normalised Difference Vegetation Index (NDVI), Soil Plant Analysis Development Chlorophyll Meter Reading (SCMR), Leaf Senescence Rate (LSR), chlorophyll fluorescence (Fv/Fm), canopy temperature (CT), phenological traits, morphological traits, yield and its attributes; and characterised into three categories viz., slow senescing, intermediate senescing and fast senescing, based on their rate of senescence. Key results Results indicate that slow-senescing genotypes had a significantly higher NDVI, SCMR and yield as compared to fast-senescing genotypes. NDVI, SCMR, Fv/Fm, days to heading, days to anthesis, days to maturity, grain weight per spike, 100-grain weight and biomass were significantly positively correlated whereas LSR and CT were significantly negatively correlated with grain yield. From the principal component analysis studies, it was revealed that genotypes with a high SCMR, a low LSR, and a high grain yield were placed together as SG genotypes. Conclusions This study confirms that genotypes with SG traits performed better under heat stress. Implications From the wild, genotypes HTW 11 (W), HTW 67 (W) and HTW 6 (W) can be utilised for heat tolerance breedings.

Novel sources of drought tolerance in sorghum landraces revealed via the analyses of genotype-by-environment interactions.

Globally, sorghum is the fifth most important crop, which is used for food, feed and fuel. However, its production and productivity are severely limited by various stresses, including drought. Hence, this study aimed to determine the responses of different drought-tolerance related traits in the Ethiopian sorghum germplasm through multi-environment field trials, thereby identifying novel sources of germplasm that can be used for breeding the crop for drought-tolerance. Three hundred twenty sorghum landraces and four improved varieties were grown at three sites within drought-prone areas (Melkassa, Mieso and Mehoni) in Ethiopia. The targeted traits were chlorophyll content at flowering (CHLF), chlorophyll content at maturity (CHLM), green leaf number at flowering (GLNF), stay-green (SG), flag leaf area (FLA), peduncle length (PDL), and panicle exertion (PAE). Multi-variate analyses of the collected data revealed the presence of high phenotypic variation in all traits. The combined and AMMI Analysis of variance showed that phenotypic variation due to the genotypes was higher for SG, CHLM, CHLF and GLNF and lower for FLA, PE and PDL in comparison with variation due to the environments or genotype by environment interactions. High broad sense heritability was observed for CHLF, CHLM, SG, GLNF, FLA, and PDL, whereas PAE showed moderate heritability. Due to the high heritability of chlorophyll content and the relatively small effect of environmental factors on it, it could serve as a criterion for selecting desirable genotypes for drought-tolerant breeding in sorghum. It has been found that chlorophyll content has a significant positive correlation with stay-green and grain yield, indicating that high chlorophyll content contributes to increasing grain yield by delaying the process of leaf senescence. The analyses of AMMI, GGE biplot, and genotype selection index revealed that several sorghum landraces outperformed the improved varieties with respect to CHLF, CHLM, and SG. Such landraces could serve as novel sources of germplasm for improving drought tolerance through breeding.

Molecular and Physiological Evaluation of Bread Wheat (Triticum aestivum L.) Genotypes for Stay Green under Drought Stress.

Water availability is considered as the main limiting factor of wheat growth illuminating the need of cultivars best adapted to drought situations for better wheat production and yield. Among these, the stay-green trait is thought to be related to the ability of wheat plants to maintain photosynthesis and CO2 assimilation, and a detailed molecular understanding of this trait may help in the selection of high-yielding, drought-tolerant wheats. The current study, therefore, evaluated the physiological responses of the selected wheat genotypes under pot-induced water stress conditions through different field capacities. The study also focused on exploring the molecular mechanisms involved in drought tolerance conferred due to the stay-green trait by studying the expression pattern of the selected PSI-associated light-harvesting complex I (LHC1) and PSII-associated LHCII gene families related to pigment-binding proteins. The results revealed that the studied traits, including relative water content, membrane stability index and chlorophyll, were variably and negatively affected, while the proline content was positively enhanced in the studied wheats under water stress treatments. Molecular diagnosis of the selected wheat genotypes using the expression profile of 06 genes, viz. TaLhca1, TaLhca2, TaLhca3, TaLhcb1, TaLhcb4 and TaLhcb6 that encodes for the LHCI and LHCII proteins, indicated variable responses to different levels of drought stress. The results obtained showed the relation between the genotypes and the severity of the drought stress condition. Among the studied genotypes, Chirya-1 and SD-28 performed well with a higher level of gene expression under drought stress conditions and may be used in genetic crosses to enrich the genetic background of common wheat against drought stress.

Canopy spectral reflectance indices correlate with yield traits variability in bread wheat genotypes under drought stress.

Drought stress is a major issue impacting wheat growth and yield worldwide, and it is getting worse as the world's climate changes. Thus, selection for drought-adaptive traits and drought-tolerant genotypes are essential components in wheat breeding programs. The goal of this study was to explore how spectral reflectance indices (SRIs) and yield traits in wheat genotypes changed in irrigated and water-limited environments. In two wheat-growing seasons, we evaluated 56 preselected wheat genotypes for SRIs, stay green (SG), canopy temperature depression (CTD), biological yield (BY), grain yield (GY), and yield contributing traits under control and drought stress, and the SRIs and yield traits exhibited higher heritability (H2) across the growing years. Diverse SRIs associated with SG, pigment content, hydration status, and aboveground biomass demonstrated a consistent response to drought and a strong association with GY. Under drought stress, GY had stronger phenotypic correlations with SG, CTD, and yield components than in control conditions. Three primary clusters emerged from the hierarchical cluster analysis, with cluster I (15 genotypes) showing minimal changes in SRIs and yield traits, indicating a relatively higher level of drought tolerance than clusters II (26 genotypes) and III (15 genotypes). The genotypes were appropriately assigned to distinct clusters, and linear discriminant analysis (LDA) demonstrated that the clusters differed significantly. It was found that the top five components explained 73% of the variation in traits in the principal component analysis, and that vegetation and water-based indices, as well as yield traits, were the most important factors in explaining genotypic drought tolerance variation. Based on the current study's findings, it can be concluded that proximal canopy reflectance sensing could be used to screen wheat genotypes for drought tolerance in water-starved environments.

STAY GREEN GENES FRAGMENT HOMOLOGY ANALYSIS OF INDONESIAN SORGHUM

Stay-green (SGR) is an essential trait in sorghum associated with grain yield under drought and high-temperature stress conditions. The study sought to analyze the homology of the stay-green gene fragment in Indonesian sorghum cultivars, comparing it with the SGR gene sequences in the GenBank database. Two primer pairs, designated as SGR_1 and SGR_2, got designed from the SGR gene of Sorghum bicolor and used to amplify seven sorghum genotypes. The DNA fragments of 300 bp and 1000 bp produced by SGR_1 and SGR_2, respectively, underwent sequencing. Amino acid analysis of the seven sorghum genotypes resulted in high homology with senescence-inducible chloroplast SGR-protein 1 from Zea mays and SGR-chloroplastic of Setaria italica. An observation on a conservative region with the SGR domain noticed SGR genes, derived from sorghum genotypes, clustered separately with those from other SGR genes available in the GenBank database. The first group consisted of a sorghum genotype (Samurai 2), the second group consisted of Super 2, Numbu, and Kawali, while the third group consisted of PI-150-20-A, Pahat, and B69 with coefficient similarity of 10%, 14%, and 30%, respectively. Although Indonesian sorghum has a different group in GenBank, it has similar nucleotide and amino acid sequences, with identity values of 95%-100% and 51%-100%, respectively. The amino acid diversity of the DNA fragments of the SGR gene is highly potential to develop molecular markers, especially the stay green character. The finding will support a sorghum breeding efficiently and precisely, especially for yield improvement under drought-stress conditions.

Crystal structure and reaction mechanism of a bacterial Mg-dechelatase homolog from the Chloroflexi Anaerolineae.

Chlorophyll degradation plays a myriad of physiological roles in photosynthetic organisms, including acclimation to light environment and nutrient remobilization during senescence. Mg extraction from chlorophyll a is the first and committed step of the chlorophyll degradation pathway. This reaction is catalyzed by the Mg-dechelatase enzyme encoded by Stay-Green (SGR). The reaction mechanism of SGR protein remains elusive since metal ion extraction from organic molecules is not a common enzymatic reaction. Additionally, experimentally derived structural information about SGR or its homologs has not yet been reported. In this study, the crystal structure of the SGR homolog from Anaerolineae bacterium was determined using the molecular replacement method at 1.85 Å resolution. Our previous study showed that three residues-H32, D34, and D62 are essential for the catalytic activity of the enzyme. Biochemical analysis involving mutants of D34 residue further strengthened its importance in the functioning of the dechelatase. Docking simulation also revealed the interaction between the D34 side chain and central Mg ion of chlorophyll a. Structural analysis showed the arrangement of D34/H32/D62 in the form of a catalytic triad that is generally found in hydrolases. The probable reaction mechanism suggests that deprotonated D34 side chain coordinates and destabilizes Mg, resulting in Mg extraction. Besides, H32 possibly acts as a general base catalyst and D62 facilitates H32 to be a better proton acceptor. Taken together, the reaction mechanism of SGR partially mirrors the one observed in hydrolases.

STAY-GREEN (SGR) genes in tomato (Solanum lycopersicum): genome-wide identification, and expression analyses reveal their involvements in ripening and salinity stress responses

STAY-GREEN (SGR) genes in tomato (Solanum lycopersicum): genome-wide identification, and expression analyses reveal their involvements in ripening and salinity stress responses