Important Agronomic Traits Research Articles

Rapid Mining of Candidate Genes for the Branchless Phenotype in Watermelon by Bulked Segregant Analysis Using Whole-genome Resequencing

Lateral branching is an important agronomic trait in horticulture plants. The aim of this project was to reveal the genetic mechanism, map the gene localization, and predict candidate genes of watermelon lateral branching. An F2 segregating population was derived from a cross between the multibranched maternal inbred line M6 and the branchless paternal inbred line N7. Two DNA pools were constructed using 20 multibranched plants and 20 branchless plants from the F2 population. Whole-genome resequencing was performed for the DNA pools (25×) and the parents (30×) to identify the genomic region associated with lateral branching. Candidate genes were predicted based on the gene annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses; then, quantitative validation of these genes was performed. The results showed that the clean reads of four samples yielded 64,295,076 to 81,658,958 bp, with sufficient genome coverage and high quality. Based on single-nucleotide polymorphism and insertions/deletions association analyses, the candidate genes were mapped to a 2.01-Mb region on chromosome 4 (22,958,925–24,971,894 bp) containing 182 annotated genes. During the KEGG and GO enrichment analyses, these genes were annotated to 10 cellular components, 10 molecular functions, and 12 biological processes. Eight candidate genes responsible for the branchless phenotype in watermelon were identified: Cla97C04G076340, Cla97C04G075820, Cla97C04G076060, Cla97C04G076250, Cla97C04G076280, Cla97C04G076380, Cla97C04G076830, and Cla97C04G075950. These genes were involved in amino acid biosynthesis and catabolism, TCP transcription factor activity, and regulation of flower development. This study offers valuable insights into the molecular mechanisms governing the branchless phenotype in watermelon. These candidate genes serve as potential targets for gene cloning and marker-assisted selection of watermelon cultivars without lateral branches.

Genetic trends in the Kenya Highland Maize Breeding Program between 1999 and 2020.

Optimization of a breeding program requires assessing and quantifying empirical genetic trends made through past efforts relative to the current breeding strategies, germplasm, technologies, and policy. To establish the genetic trends in the Kenyan Highland Maize Breeding Program (KHMP), a two-decade (1999-2020) historical dataset from the Preliminary Variety Trials (PVT) and Advanced Variety Trials (AVT) was analyzed. A mixed model analysis was used to compute the genetic gains for traits based on the best linear unbiased estimates in the PVT and AVT evaluation stages. A positive significant genetic gain estimate for grain yield of 88 kg ha-1 year-1 (1.94% year-1) and 26 kg ha-1 year-1 (0.42% year-1) was recorded for PVT and AVT, respectively. Root lodging, an important agronomic trait in the Kenya highlands, had a desired genetic gain of -2.65% year-1 for AVT. Results showed improvement in resistance to Turcicum Leaf Blight (TLB) with -1.19% and -0.27% year-1 for the PVT and AVT, respectively. Similarly, a significant genetic trend of -0.81% was noted for resistance to Gray Leaf Spot (GLS) in AVT. These findings highlight the good progress made by KHMP in developing adapted maize hybrids for Kenya's highland agroecology. Nevertheless, the study identified significant opportunities for the KHMP to make even greater genetic gains for key traits with introgression of favorable alleles for various traits, implementing a continuous improvement plan including marker-assisted forward breeding, sparse testing, and genomic selection, and doubled haploid technology for line development.

Overexpression of CsBRC, an F-box gene from Camellia sinensis, increased the plant branching in tobacco and rice.

Tea plant (Camellia sinensis [L.]) is one of the most important crops in China, and tea branch is an important agronomic trait that determines the yield of tea plant. In previous work focused on GWAS that detecting GWAS signals related to plant architecture through whole genome re-sequencing of ancient tea plants, a gene locus TEA 029928 significantly related to plant type was found. Sequence alignment results showed that this gene belonged to the F-box family. We named it CsBRC. CsBRC-GFP fusion proteins were mainly localized in the plasma membrane. By comparing the phenotypes of CsBRC transgenic tobacco and WT tobacco, it was found that the number of branches of transgenic tobacco was significantly higher than that of wild-type tobacco. Through RNA-seq analysis, it was found that CsBRC affects the branching development of plants by regulating the expression of genes related to brassinosteroid synthesis pathway in plants. In addition, overexpression of CsBRC in rice could increase tiller number, grain length and width, and 1,000-grain weight.

Quantitative Trait Loci Sequencing and Genetic Mapping Reveal Two Main Regulatory Genes for Stem Color in Wax Gourds.

Stem color is an important agronomic trait of wax gourds. However, its regulatory genes have not been identified. In this study, 105 inbred lines constructed from two parents (GX-71 and MY-1) were sequenced and quantitative trait loci sequencing was used to mine the genes that regulate stem color in wax gourds. The results identified two quantitative trait loci related to stem color, qSC5 and qSC12, located on Chr05 (11,134,567-16,459,268) and Chr12 (74,618,168-75,712,335), respectively. The qSC5 had a phenotypic variation rate of 36.9% and a maximum limit of detection of 16.9. And the qSC12 had a phenotypic variation rate of 20.9%, and a maximum limit of detection of 11.2. Bch05G003950 (named BchAPRR2) and Bch12G020400 were identified as candidate genes involved in stem color regulation in wax gourds. The chlorophyll content and expression of BchAPRR2 and Bch12G020400 were significantly higher in green-stemmed wax gourds than in white-stemmed ones. Therefore, BchAPRR2 and Bch12G020400 were considered the main and secondary regulatory genes for wax gourd stem color, respectively. Finally, InDel markers closely linked to BchAPRR2 were developed to validate the prediction of wax gourd stem color traits in 55 germplasm lines, with an accuracy of 81.8%. These findings lay the foundation for exploring the genetic regulation of wax gourd stem color and future research on wax gourd breeding.

Genome-Wide Association Study and Transcriptome Analysis Provide Candidate Genes for Agronomic Traits of Agaricus bisporus

Agaricus bisporus, belonging to the genus Agaricus and the family Agaricaceae, is a popular commercially cultivated mushroom with rich nutritional and medical values. Cultivation of A. bisporus requires superior cultivars. Understanding the differences between wild and cultivated accession at the genetic level is crucial for mining loci and genes associated with cultivation-related traits, informing future breeding directions. Additionally, the identification of loci and genes associated with important agronomic traits (e.g., yield and quality) facilitates mushroom molecular breeding. In this study, we conducted sequencing of 200 strains of A. bisporus and analyzed genomic variations. Population structure and genetic relationships were investigated for 200 strains. Selection signs and genes were also obtained after selection sweep analysis. Thirteen candidate genes in the selective elimination regions had significantly different expression patterns at the fruit body stage. Moreover, six genes were identified for the esterase isozyme type through a combination of GWAS and RNA data. This study provides insight into environmental adaptation at the genetic base, providing valuable genetic resources for button mushroom molecular breeding to improve environmental adaptation, quality, or yield.

Identification and Fine Mapping of Quantitative Trait Loci for Tiller Angle Using Chromosome Segment Substitution Lines in Rice (Oryza Sativa L.)

The tiller angle, which is an important agronomic trait, determines plant architecture and greatly influences the grain yield of rice. In this study, a population of chromosome segment substitution lines derived from a cross between a japonica variety with a compact plant architecture—Koshihikari—and an indica variety with a spread-out plant architecture—Nona Bokra—was used to investigate the genetic basis of the tiller angle. Five quantitative trait loci (qTA1, qTA5, qTA9-1, qTA9-2, and qTA11) for the tiller angle were detected on chromosomes 1, 5, 9, 9, and 11 in two different environments. The phenotypic variation in these QTLs ranged from 3.78% to 8.22%. Two pairs of digenic epistatic QTLs were detected in Lingshui. The epistatic interaction explained 15.19% and 13.60% of the phenotypic variance, respectively. Among the five QTLs, qTA9-2 was detected in both environments. An F2 mapping population containing the qTA9-2 QTL was established. The location of qTA9-2 was narrowed down to a 187 kb region between InDel markers M9 and M10 on chromosome 9. Thirty open reading frames (ORFs), including TAC1, a gene known to regulate the tiller angle, were identified in this region. The gene sequencing results suggested that a base substitution from G to A at position 1557 in the 3′-untranslated region led to a difference in the expression of qTA9-2 in Koshihikari and Nona Bokra. These findings provide a potential gene resource for the improvement of rice plant architecture.

Population-level exploration of alternative splicing and its unique role in controlling agronomic traits of rice.

Alternative splicing (AS) plays crucial roles in regulating various biological processes in plants. However, the genetic mechanisms underlying AS and its role in controlling important agronomic traits in rice (Oryza sativa) remain poorly understood. In this study, we explored AS in rice leaves and panicles using the rice minicore collection. Our analysis revealed a high level of transcript isoform diversity, with approximately one-fifth of the potential isoforms acting as major transcripts in both tissues. Regarding the genetic mechanism of AS, we found that the splicing of 833 genes in the leaf and 1,230 genes in the panicle was affected by cis-genetic variation. Twenty-one percent of these AS events could only be explained by large structural variations. Approximately 77.5% of genes with significant splicing quantitative trait loci (sGenes) exhibited tissue-specific regulation, and AS can cause 26.9% (leaf) and 23.6% (panicle) of sGenes to have altered, lost, or gained functional domains. Additionally, through splicing-phenotype association analysis, we identified phosphate-starvation-induced RING-type E3 ligase (OsPIE1; LOC_Os01g72480), whose splicing ratio was significantly associated with plant height. In summary, this study provides an understanding of AS in rice and its contribution to the regulation of important agronomic traits.

SSR-BASED ASSESSMENT OF GENETIC DIVERSITY IN TOMATO CULTIVARS AND LINES GROWN IN KAZAKHSTAN

Tomato (Solanum lycopersicum L.) is a versatile crop known for its nutritional value and health benefits, thriving in diverse climates worldwide. Nevertheless, regional variations persist in tomato yield, with Kazakhstan serving as an example of lower productivity in contrast to global averages. Closing this disparity requires comprehensive genetic studies and breeding efforts. This study is focused on the genetic diversity of 49 tomato cultivars and hybrids sourced from Kazakh Research Institute of Fruit and Vegetable Growing (Almaty), employing 10 SSR markers associated with important agronomic traits. SSR genotyping unveiled polymorphisms across 6 markers with variable allele numbers. Genetic diversity metrics highlighted significant genetic diversity within both outdoor and greenhouse tomato cultivars and lines. Bayesian clustering, Neighbor-joining (NJ) clustering, and principal coordinate analysis (PCoA) delineated genetic differentiation between outdoor and greenhouse tomatoes with small admixture, indicating distinct breeding directions for these two types. Highly polymorphic SSRs (PIC > 0.5) associated with essential fruit traits emerge as promising targets for marker-assisted selection (MAS) that can be used to enhance tomato breeding efficiency in Kazakhstan. According to 8 SSRs, 22 out of 30 outdoor accessions and 8 greenhouse tomato accessions were genetically uniform. This study offers comprehensive insights into the genetic diversity and population structure of tomato cultivars and lines in Kazakhstan, laying the foundation for informed breeding endeavors aimed at bolstering yield and resilience in tomato crops.

Photosynthesis regulates tillering bud elongation and nitrogen-use efficiency via sugar-induced NGR5 in rice.

Rice tillering is one of the most important agronomical traits largely determining grain yield. Photosynthesis and nitrogen availability are two important factors affecting rice tiller bud elongation; however, underlying mechanism and their cross-talk is poorly understood. Here, we used map-based cloning, transcriptome profiling, phenotypic analysis, and molecular genetics to understand the roles of the Decreased Tiller Number 1 (DTN1) gene that encodes the fructose-1,6-bisphosphate aldolase and involvesin photosynthesis required for light-induced axillary bud elongation in rice. Deficiency of DTN1 results in the reduced photosynthetic rate and decreased contents of sucrose and other sugars in both leaves and axillary buds, and the reduced tiller number in dtn1 mutant could be partially rescued by exogenous sucrose treatment. Furthermore, we found that the expression of nitrogen-mediated tiller growth response 5 (NGR5) was remarkably decreased in shoot base of dtn1-2, which can be activated by sucrose treatment. Overexpression of NGR5 in the dtn1-2 could partially rescue the reduced tiller number, and the tiller number of dtn1-2 was insensitive to nitrogen supply. This work demonstrated that the sugar level regulated by photosynthesis and DTN1 could positively regulate NGR5 expression, which coordinates the cross-talk between carbon and nitrate to control tiller bud outgrowth in rice.

Identification of the function of a key gene NnHCT1 in lignin synthesis in petioles of Nelumbo nucifera

Leaf petiole or stem strength is an important agronomic trait affecting the growth of underground organs as a channel for material exchange and plays a vital role in the quality and yield of crops and vegetables. There are two different types of petioles in lotus, floating leaf petioles and vertical leaf petioles; however, the internal difference mechanism between these petioles is unclear. In this study, we investigated the differences between the initial vertical leaf petioles and the initial floating leaf petioles based on RNA sequencing (RNA-seq), and >2858 differentially expressed genes were annotated. These genes were chiefly enriched in phenylpropanoid biosynthesis, which is the source of the lignin and cellulose in petioles and stems. Lignin biology-related gene NnHCT1 was identified, and subsequent biological function validation demonstrated that the transient overexpression of NnHCT1 significantly increased the lignin and cellulose contents in lotus petioles and tobacco leaves. In contrast, silencing NnHCT1 through virus-induced gene silencing significantly reduced petiole lignin synthesis. Additionally, differentially up-regulated MYB family transcription factors were identified using RNA-seq. Yeast-one-hybrid and dual-luciferase reporter assays demonstrated that MYB4 could bind to the NnHCT1 promoter and up-regulate NnHCT1 expression. These findings demonstrate the significant potential of NnHCT1 to enhance lignin synthesis, thereby improving stem or petiole resistance to stunting and explaining the need for the study of differential petiole relationships in plants.

Fine mapping and transcriptome profiling reveal CpAPRR2 to modulate immature fruit rind color formation in zucchini (Cucurbita pepo).

A large fragment deletion of CpAPRR2, encoding a two-component response regulator-like protein, which influences immature white rind color formation in zucchini (Cucurbita pepo). Fruit rind color is an important agronomic trait that affects commodity quality and consumer choice in zucchini (Cucurbita pepo). However, the molecular mechanism controlling rind color is unclear. We characterized two zucchini inbred lines: '19' (dark green rind) and '113' (white rind). Genetic analysis revealed white immature fruit rind color to be controlled by a dominant locus (CpW). Combining bulked segregant analysis sequencing (BSA-seq) and Kompetitive Allele-Specific PCR (KASP) markers, we mapped the CpW locus to a 100.4kb region on chromosome 5 and then narrow down the candidate region to 37.5kb using linkage analysis of 532 BC1 and 1613 F2 individuals, including 6 coding genes. Among them, Cp4.1LG05g02070 (CpAPRR2), encoding a two-component response regulator-like protein, was regarded to be a promising candidate gene. The expression level of CpAPRR2 in dark green rind was significantly higher than that in white rind and was induced by light. A deletion of 2227bp at the 5' end of CpAPRR2 in '113' might explain the white phenotype. Further analysis of allelic diversity in zucchini germplasm resources revealed rind color to be associated with the deletion of CpAPRR2. Subcellular localization analysis indicated that CpAPRR2 was a nuclear protein. Transcriptome analysis using near-isogenic lines with dark green (DG) and white (W) rind indicated that genes involved in photosynthesis and porphyrin metabolism pathways were enriched in DG compared with W. Additionally, chlorophyll synthesis-related genes were upregulated in DG. These results identify mechanisms of zucchini rind color and provide genetic resources for breeding.

The lncRNA1-miR6288b-3p-PpTCP4-PpD2 module regulates peach branch number by affecting brassinosteroid biosynthesis.

Branch number is one of the most important agronomic traits of fruit trees such as peach. Little is known about how LncRNA and/or miRNA modules regulate branching through transcription factors. Here, we used molecular and genetic tools to clarify the molecular mechanisms underlying brassinosteroid (BR) altering plant branching. We found that the number of sylleptic branch and BR content in pillar peach ('Zhaoshouhong') was lower than those of standard type ('Okubo'), and exogenous BR application could significantly promote branching. PpTCP4 expressed great differentially comparing 'Zhaoshouhong' with 'Okubo'. PpTCP4 could directly bind to DWARF2 (PpD2) and inhibited its expression. PpD2 was the only one differentially expressed key gene in the path of BR biosynthesis. At the same time, PpTCP4 was identified as a target of miR6288b-3p. LncRNA1 could act as the endogenous target mimic of miR6288b-3p and repress expression of miR6288b-3p. Three deletions and five SNP sites of lncRNA1 promoter were found in 'Zhaoshouhong', which was an important cause of different mRNA level of PpTCP4 and BR content. Moreover, overexpressed PpTCP4 significantly inhibited branching. A novel mechanism in which the lncRNA1-miR6288b-3p-PpTCP4-PpD2 module regulates peach branching number was proposed.

A genome-wide association study reveals novel loci and candidate genes associated with plant height variation in Medicago sativa

BackgroundPlant height (PH) is an important agronomic trait influenced by a complex genetic network. However, the genetic basis for the variation in PH in Medicago sativa remains largely unknown. In this study, a comprehensive genome-wide association analysis was performed to identify genomic regions associated with PH using a diverse panel of 220 accessions of M. sativa worldwide.ResultsOur study identified eight novel single nucleotide polymorphisms (SNPs) significantly associated with PH evaluated in five environments, explaining 8.59–12.27% of the phenotypic variance. Among these SNPs, the favorable genotype of chr6__31716285 had a low frequency of 16.4%. Msa0882400, located proximal to this SNP, was annotated as phosphate transporter 3;1, and its role in regulating alfalfa PH was supported by transcriptome and candidate gene association analysis. In addition, 21 candidate genes were annotated within the associated regions that are involved in various biological processes related to plant growth and development.ConclusionsOur findings provide new molecular markers for marker-assisted selection in M. sativa breeding programs. Furthermore, this study enhances our understanding of the underlying genetic and molecular mechanisms governing PH variations in M. sativa.

Genome-editing of a circadian clock gene TaPRR95 facilitates wheat peduncle growth and heading date

Plant height and heading date are important agronomic traits in wheat (Triticum aestivum L.) that affect final grain yield. In wheat, knowledge of pseudo-response regulator (PRR) genes on agronomic traits is limited. Here, we identify a wheat TaPRR95 gene by genome-wide association study to be associated with plant height. Triple allele mutant plants produced by CRISPR/Cas9 show increased plant height, particularly the peduncle, with an earlier heading date. The longer peduncle is mainly caused by the increased cell elongation at its upper section, whilst the early heading date is accompanied by elevated expression of flowering genes, such as TaFT and TaCO1. A peduncle-specific transcriptome analysis reveals up-regulated photosynthesis genes and down-regulated IAA/Aux genes for auxin signaling in prr95aabbdd plants that may act as a regulatory mechanism to promote robust plant growth. A haplotype analysis identifies a TaPRR95-B haplotype (Hap2) to be closely associated with reduced plant height and increased thousand-grain weight. Moreover, the Hap2 frequency is higher in cultivars than that in landraces, suggesting the artificial selection on the allele during wheat breeding. These findings suggest that TaPRR95 is a regulator for plant height and heading date, thereby providing an important target for wheat yield improvement.

Identification and Characterization of HS4-Mediated Hybrid Seed Shattering in Rice

Seed shattering is an adaptive feature of seed dispersal in wild rice, and it is also an important agronomic trait affecting yield. Reduced seed shattering was a significant progress during rice domestication. However, the evolutionary pathway and molecular mechanism of hybrid seed shattering remain largely unknown. In order to gain a deeper understanding of the regulation of hybrid seed shattering, HS4, a locus conferring hybrid seed shattering between Oryza sativa and Oryza glaberrima, was identified and fine mapped to a 13.5-kb genomic region containing two putative genes during the development of chromosomal segment substitution lines (CSSLs). Expression analysis indicated that the hybrid seed shattering was not related to the expression of HS4. Preliminary research on the molecular mechanism of HS4-mediated hybrid seed shattering indicated that HS4HJX74 and HS4HP61 may form a multimer in heterozygotes, achieving the original function of a trihelix transcription factor through protein interaction. The identification and characterization of HS4 in this study not only provides new insights into the molecular mechanisms underlying hybrid seed shattering, but also provides a potential target for genome editing to reduce the difficulty of hybridization between the two species, facilitating hybrid breeding and increasing yield in rice.

Fine-mapping and candidate gene analysis of tuber eye depth in potato

Eye depth is an important agronomic trait affecting tubers' appearance, quality, and processing suitability. Hence, cultivating varieties with uniform shapes and shallow eye depth are important goals for potato breeding. In this study, based on the primary mapping of the tuber eye-depth locus using a small primary-segregating population, a large secondary-segregating population with 2,100 individuals was used to map the eye-depth locus further. A major quantitative trait locus for eye-depth on chromosome 10 was identified (designated qEyd10.1) using BSA-seq and traditional QTL mapping method. The qEyd10.1 could explain 55.0% of the eye depth phenotypic variation and was further narrowed to a 309.10 kb interval using recombinant analysis. To predict candidate genes, tissue sectioning and RNA-seq of the specific tuber tissues were performed. Genes encoding members of the peroxidase superfamily with likely roles in indole acetic acid regulation were considered the most promising candidates. These results will facilitate marker-assisted selection for the shallow-eye trait in potato breeding and provide a solid basis for eye-depth gene cloning and the analysis of tuber eye-depth regulatory mechanisms.

Quantitative Trait Locus Mapping for Plant Height and Branch Number in CCRI70 Recombinant Inbred Line Population of Upland Cotton (Gossypium hirsutum).

Upland cotton accounts for a high percentage (95%) of the world's cotton production. Plant height (PH) and branch number (BN) are two important agronomic traits that have an impact on improving the level of cotton mechanical harvesting and cotton yield. In this research, a recombinant inbred line (RIL) population with 250 lines developed from the variety CCRI70 was used for constructing a high-density genetic map and identification of quantitative trait locus (QTL). The results showed that the map harbored 8298 single nucleotide polymorphism (SNP) markers, spanning a total distance of 4876.70 centimorgans (cMs). A total of 69 QTLs for PH (9 stable) and 63 for BN (11 stable) were identified and only one for PH was reported in previous studies. The QTLs for PH and BN harbored 495 and 446 genes, respectively. Combining the annotation information, expression patterns and previous studies of these genes, six genes could be considered as potential candidate genes for PH and BN. The results could be helpful for cotton researchers to better understand the genetic mechanism of PH and BN development, as well as provide valuable genetic resources for cotton breeders to manipulate cotton plant architecture to meet future demands.

Physical Seed Dormancy in Legumes: Molecular Advances and Perspectives.

Physical dormancy of seeds is a form of dormancy due to the presence of an impermeable seed coat layer, and it represents a feature for plants to adapt to environmental changes over an extended period of phylogenetic evolution. However, in agricultural practice, physical dormancy is problematic. because it prevents timely and uniform seed germination. Therefore, physical dormancy is an important agronomical trait to target in breeding and domestication, especially for many leguminous crops. Compared to the well-characterized physiological dormancy, research progress on physical dormancy at the molecular level has been limited until recent years, due to the lack of suitable research materials. This review focuses on the structure of seed coat, factors affecting physical dormancy, genes controlling physical dormancy, and plants suitable for studying physical dormancy at the molecular level. Our goal is to provide a plethora of information for further molecular research on physical dormancy.

GWAS analysis reveals the genetic basis of blast resistance associated with heading date in rice.

Rice blast is a destructive fungal disease affecting rice plants at various growth stages, significantly threatening global yield stability. Development of resistant rice cultivars stands as a practical means of disease control. Generally, association mapping with a diversity panel powerfully identifies new alleles controlling trait of interest. On the other hand, utilization of a breeding panel has its advantage that can be directly applied in a breeding program. In this study, we conducted a genome-wide association study (GWAS) for blast resistance using 296 commercial rice cultivars with low population structure but large phenotypic diversity. We attempt to answer the genetic basis behind rice blast resistance among early maturing cultivars by subdividing the population based on its Heading date 1 (Hd1) functionality. Subpopulation-specific GWAS using the mixed linear model (MLM) based on blast nursery screening conducted in three years revealed a total of 26 significant signals, including three nucleotide-binding site leucine-rich repeat (NBS-LRR) genes (Os06g0286500, Os06g0286700, and Os06g0287500) located at Piz locus on chromosome 6, and one at the Pi-ta locus (Os12g0281300) on chromosome 12. Haplotype analysis revealed blast resistance associated with Piz locus was exclusively specific to Type 14 hd1 among japonica rice. Our findings provide valuable insights for breeding blast resistant rice and highlight the applicability of our elite cultivar panel to detect superior alleles associated with important agronomic traits.

MicroRNA regulates cytokinin induced parthenocarpy in cucumber (Cucumis sativus L.)

Parthenocarpy is one of the most important agronomic traits for fruit yield in cucumbers. However, the precise gene regulation and the posttranscriptional mechanism are elusive. In the presented study, one parthenocarpic line DDX and non-parthenocarpic line ZK were applied to identify the microRNAs (miRNAs) involved in parthenocarpic fruit formation. The differential expressed miRNAs among parthenocarpic fruit of forchlorfenuron (CPPU) treated ZK (ZK-CPPU), pollinated ZK (ZK-P), non-pollinated DDX (DDX-NP) were compared with the non-parthenocarpic fruits of non-pollinated ZK (ZK-NP). It indicated 98 miRNAs exhibited differential expression were identified. Notably, a significant proportion of these miRNAs were enriched in the signal transduction pathway of plant hormones, as identified by the KEGG pathway analysis. qRT-PCR validation indicated that CsmiR156 family was upregulated in the ZK-NP while downregulated in ZK-CPPU, ZK-P, and DDX-NP at 1 day after anthesis. Meanwhile, the opposite trend was observed for CsmiR164a. In ZK-CPPU, ZK-P, and DDX-NP, CsmiRNA156 genes (CsSPL16 and CsARR9-like) were upregulated while CsmiRNA164a genes (CsNAC6, CsCUC1, and CsNAC100) were downregulated. The GUS and dual luciferase assay validated that CsmiR156a inhibited while CsmiR164a induced their target genes' transcription. This study presents novel insights into the involvement of CsmiR156a and CsmiR164a in the CK-mediated posttranscriptional regulation of cucumber parthenocarpy, which will aid future breeding programs.