The ABORTED MICROSPORES (AMS) gene is crucial for tapetal cell development and pollen formation, but its role in Upland cotton (Gossypium hirsutum) has not been previously documented. This study identified GhAMS11 as a key transcription factor, with its high expression specifically observed during the S4-S6 stages of anther development, a critical period for tapetal activity and pollen formation. Subcellular localization confirmed that GhAMS11 was located in the nucleus. CRISPR/Cas9 knockout of GhAMS11 resulted in pollen inviability, with mutants displaying abnormal tapetal development and defective pollen exine formation. TUNEL assays highlighted GhAMS11's involvement in proper tapetal programmed cell death (PCD). Additionally, GhAMS11 was found to activate GhMS188 expression, as demonstrated by dual-luciferase assays and EMSA assays, with their interaction confirmed through LCI assays, yeast two-hybrid assays and GST pull down assays. Deletion of GhMS188 led to pollen sterility, grain collapse, and impaired pollen exine formation. Thus, this research identified the bHLH transcription factor GhAMS11, addressing a gap in AMS gene research in Upland cotton, and elucidated its key regulatory role in pollen development in cooperation with GhMS188.

IntroductionVerticillium wilt, incited by the soilborne fungus Verticillium dahliae, is a severe threat to global cotton (Gossypium spp.) production, resulting in significant yield losses and reduced fiber quality.MethodsTo uncover the genetic and molecular basis of resistance to this devastating disease, we combined genome-wide association study (GWAS) and transcriptomic analyses in a natural population of 355 upland cotton accessions.ResultsGWAS identified a stable major-effect quantitative trait locus (QTL), qVW-A01-2, on chromosome A01, which harbors the candidate gene GhAMT2, encoding a high-affinity ammonium transporter. Transcriptomic profiling revealed that GhAMT2 was significantly upregulated at 12 hours post-inoculation with V. dahliae, coinciding with the activation of immune signaling pathways. Weighted Gene Co-expression Network Analysis (WGCNA) further linked GhAMT2 to critical defense pathways, including lignin biosynthesis, salicylic acid signaling, and reactive oxygen species (ROS) homeostasis, suggesting its role in cell wall reinforcement and systemic immune responses. Functional validation through virus-induced gene silencing (VIGS) confirmed that silencing GhAMT2 compromised disease resistance. In contrast, transgenic Arabidopsis plants overexpressing GhAMT2 exhibited enhanced resistance to V. dahliae, demonstrating its essential role in defense regulation.DiscussionThese findings establish GhAMT2 as a key regulator of cotton resistance to Verticillium wilt and highlight its potential for marker-assisted breeding and genetic engineering to improve disease-resistant cotton varieties.

Male reproductive development is fundamental to the life cycle of flowering plants, culminating in seed production. Aberrations in anther development frequently lead to male sterility, yet the underlying molecular mechanisms in upland cotton (Gossypium hirsutum) remain largely uncharacterized. The R2R3-MYB family of transcription factors are known key regulators of diverse developmental processes, including male fertility in several model species. Here, we identify and functionally characterize GhMYB35, an R2R3-MYB transcription factor that plays an essential role in cotton anther development. CRISPR/Cas9-mediated knockout of GhMYB35 resulted in complete male sterility, with mutants (ghmyb35) exhibiting indehiscent anthers, shorter filaments, and a total absence of viable pollen. Expression analyses reveal that GhMYB35 is predominantly expressed in anthers, with peak expression of its A- and D-subgenome homoeologs occurring at developmental stage 7. Subcellular localization results show that bothGhMYB35_AandGhMYB35_Dare nuclear-localized transcription factors. Furthermore, the total absence of GhMYB35 leads to pollen abortion and subsequent anther collapse without dehiscence. Collectively, our findings establish GhMYB35 as a critical regulator of anther maturation, thereby elucidating a key component of the molecular network governing male fertility in cotton.

Dense planting can create a shade environment which will stimulate plants to produce a series of shade avoidance responses (SAS). Thus far, slow progress has been made in research into the molecular mechanism of SAS in cotton. In this study, GhFPF1 was identified as a positive regulator of SAS in cotton. The expression of GhFPF1 was significantly upregulated in cotton stems and leaves in the low R:FR conditions. After mutating GhFPF1 using CRISPR technology, the elongation response of the mutant was weakened under high-density cultivating conditions. The constitutive expression of GhFPF1 in cotton resulted in typical SAS, such as a higher plant height, slow branching development and a reduced leaf inclination angle. Further assays showed that GhFPF1 interacted with GhNF-YA3, which is a transcription factor of nuclear factor Y (NF-Y) family. GhNF-YA3 was downregulated in low R:FR environment. The silencing of GhNF-YA3 led to significant internode elongation, which was similar to the phenotype caused by overexpressing GhFPF1. Comprehensive studies suggested that the interaction between GhFPF1 and GhNF-YA3 competitively weakened the transcriptional regulation of three light-harvesting chlorophyll a/b-binding protein (Lhc) genes by GhNF-YA3. In addition, GhFPF1 was proved to be the target gene of HD-Zip II transcription factors GhHB2 and GhHB4 which were considered as positive regulators of SAS in other plants. This study uncovered the molecular mechanism by which GhFPF1 regulates SAS in cotton and enriched the regulatory network of SAS in plants.

As an important part of the plant reproductive system, pollen tubes play a key role in maintaining normal male fertility. Abnormal pollen tube germination severely impacts male fertility, yet its regulatory mechanisms remain poorly understood. Ascorbic acid oxidase (AAO) plays an important role in pollen tube germination, and the functional role of AAO in male sterility of flowering plants such as Arabidopsis thaliana and barley has been reported. However, molecular cloning and functional identification of AAO in cotton remain unclear. In this study, the GhAAO9 gene, which is closely associated with male sterility in cotton, was successfully cloned and characterized. Following its knockout using CRISPR-Cas9, it was observed that the mutant pollen failed to disperse properly and exhibited a near-complete loss of viability (96 %), which is the primary cause of the male sterility phenotype. A minority of mutant pollen grains (∼4 %) that retained partial viability displayed severely impaired pollen tube germination and elongation in vitro… qRT-PCR analysis confirmed a significant reduction in GhAAO9 transcript levels in ghaao9 mutant anthers/pollen relative to the WT, which, together with the frameshift mutations, substantiates that the observed male sterility arises from deficient GhAAO9 function during pollen development. The electrophoretic mobility shift assay (EMSA) further confirmed the in vitro interaction between GhGATA1 and the regulatory region of GhAAO9. The dual-luciferase reporter assay confirmed that GhGATA1 suppresses the expression of GhAAO9, which subsequently results in the manifestation of a sterile phenotype. This study demonstrated that GhGATA1 regulates GhAAO9 expression, influencing pollen tube germination in cotton and offering valuable insights into the mechanisms underlying male sterility. While AAO functions have been characterized in Arabidopsis and cereals, cotton (Gossypium hirsutum)-an allotetraploid species with complex reproductive biology-remains unexplored. Given cotton's global agricultural importance and the unique challenges in its pollen tube guidance mechanisms, elucidating GhAAO9's role addresses a critical gap in understanding male sterility in fiber crops.

Sporopollenin, as the main component of the pollen exine, is a highly resistant polymer that provides structural integrity under unfavourable environmental conditions. Tetraketone alpha-pyrone reductase 1 (TKPR1) is essential for sporopollenin formation, catalyzing the reduction of tetraketone carbonyl to hydroxylated alpha-pyrone. The functional role of TKPR1 in male sterility has been reported in flowering plants such as maize, rice, and Arabidopsis. However, the molecular cloning and functional characterization of TKPR1 in cotton remain unaddressed. In this study, we identified 68 TKPR1s from four cotton species, categorized into three clades. Transcriptomics and RT-qPCR demonstrated that GhTKPR1_8 exhibited typical expression patterns in the tetrad stage of the anther. GhTKPR1_8 was localized to the endoplasmic reticulum. Moreover, ABORTED MICROSPORES (GhAMS) transcriptionally activated GhTKPR1_8 as indicated by luciferase complementation tests. GhTKPR1_8-knockdown inhibited anther dehiscence and reduced pollen viability in cotton. Additionally, overexpression of GhTKPR1_8 in the attkpr1 mutant restored its male sterile phenotype. This study offers novel insights into the investigation of TKPR1 in cotton while providing genetic resources for studying male sterility.

Plant phenotype assessment approaches have facilitated crop improvement in recent years by providing opportunities for the dissection of complex nature quantitative traits. Fiber quality and yield are significant economic traits with a complex nature mainly due to interactions between its genetic architecture and the surrounding environment. These economic traits showed stagnant performance in recent decades due to eroded genetic diversity via the selection process. It necessitates exploiting conserved germplasm resources to rebuild genetic diversity within cotton cultivars. Compared to the conventional univariate selection methods using a single trait at a time, the multivariate analysis extends its selection to multiple variables for the selection of genotypes. In evaluating a core collection of cotton over two years using various phenotyping methods, multivariate analysis, particularly principal component analysis (PCA), was employed to assess genetic diversity. The PCA analysis demonstrated substantial variability of 39.83% and 45.37% across two years, highlighting a significant impact on yield and fiber quality attributes. The cumulative eigenvalues of principal components exceeding 80% supported the classification of accessions into six distinct groups based on their trait variability. Both PCA and cluster analysis successfully categorized accessions into six groups, indicating high diversity and suggesting their potential utility in breeding programs. The diverse genotypes identified in this study would be potential resources for variety development and accelerated breeding programs, particularly focusing on stress resilience under climate change scenarios. By utilizing these diverse genotypes, there is an opportunity to enhance breeding strategies and accelerate the development of improved cotton varieties with enhanced yield and fiber quality attributes. The significance of our study lies in its demonstration of the importance of genetic diversity in improving yield and fiber quality attributes, thus underscoring its relevance to cotton breeding programs.

Caffeoyl coenzyme A-O-methyltransferase (CCoAOMT) has a critical function in the lignin biosynthesis pathway. However, its functions in cotton are not clear. In this research, we observed 50 CCoAOMT genes from four cotton species, including two diploids (Gossypium arboretum, 9, and Gossypium raimondii, 8) and two tetraploids (Gossypium hirsutum, 16, and Gossypium barbadense, 17), performed bioinformatic analysis, and focused on the involvement and functions of GhCCoAOMT7 in lignin synthesis of Gossypium hirsutum. CCoAOMT proteins were divided into four subgroups based on the phylogenetic tree analysis. Motif analysis revealed that all CCoAOMT proteins possess conserved Methyltransf_3 domains, and conserved structural features were identified based on the genes’ exon-intron organization. A synteny analysis suggested that segmental duplications were the primary cause in the expansion of the CCoAOMT genes family. Transcriptomic data analysis of GhCCoAOMTs revealed that GhCCoAOMT2, GhCCoAOMT7, and GhCCoAOMT14 were highly expressed in stems. Subcellular localization experiments of GhCCoAOMT2, GhCCoAOMT7, and GhCCoAOMT14 showed that GhCCoAOMT2, GhCCoAOMT7, and GhCCoAOMT14 were localized in the nucleus and plasma membrane. However, there are no cis-regulatory elements related to lignin synthesis in the GhCCoAOMT7 gene promoter. GhCCoAOMT7 expression was inhibited by virus-induced gene silencing technology to obtain gene silencing lines, the suppression of GhCCoAOMT7 expression resulted in a 56% reduction in the lignin content in cotton stems, and the phloroglucinol staining area corresponding to the xylem was significantly decreased, indicating that GhCCoAOMT7 positively regulates lignin synthesis. Our results provided fundamental information regarding CCoAOMTs and highlighted their potential functions in cotton lignin biosynthesis and lignification.

Background Neutral/alkaline invertases (N/AINVs) play a crucial role in plant growth, development, and stress response, by irreversibly hydrolyzing sucrose into glucose and fructose. However, research on cotton in this area is limited. This study aims to investigate GhN/AINV23 , a neutral/alkaline invertase in cotton, including its characteristics and biological functions. Results In our study, we analyzed the sequence information, three-dimensional (3D) model, phylogenetic tree, and cis-elements of GhN/AINV23 . The localization of GhN/AINV23 was determined to be in the cytoplasm and cell membrane. Quantitative real-time polymerase chain reaction (qRT-PCR) results showed that GhN/AINV23 expression was induced by abscisic acid (ABA), exogenous sucrose and low exogenous glucose, and inhibited by high exogenous glucose. In Arabidopsis , overexpression of GhN/AINV23 promoted vegetative phase change, root development, and drought tolerance. Additionally, the virus-induced gene silencing (VIGS) assay indicated that the inhibition of GhN/AINV23 expression made cotton more susceptible to drought stress, suggesting that GhN/AINV23 positively regulates plant drought tolerance. Conclusion Our research indicates that GhN/AINV23 plays a significant role in plant vegetative phase change, root development, and drought response. These findings provide a valuable foundation for utilizing GhN/AINV23 to improve cotton yield.

棉花是重要的经济作物之一，而棉花倒伏是导致棉花产量下降的原因之一。所以，了解棉花倒伏的原因，挖掘棉花茎秆生长关键基因，进一步利用关键基因来增强棉花茎秆机械强度进而使棉花抗倒伏是非常重要的。咖啡酰辅酶A-O-甲基转移酶（Caffeoyl-coenzyme A-O-methyltransferases, CCoAOMT）是一种参与木质素合成途径的关键酶。本研究对棉花中CCoAOMT基因家族进行系统进化分析，在亚洲棉、雷蒙德氏棉、陆地棉和海岛棉中分别鉴定到9个、8个、16个和17个CCoAOMT基因，与拟南芥中的7个CCoAOMT成员具有同源性。分析陆地棉中鉴定出来的16个基因的启动子顺式作用元件，发现GhCCoAOMT7不含有木质素相关顺式作用元件。对GhCCoAOMT7基因结构分析发现含有5个外显子和4个内含子，编码一条含378个氨基酸的蛋白序列，CDS长957bp，亚细胞定位结果显示该基因编码的蛋白位于细胞核和细胞膜。病毒诱导的基因沉默VIGS技术将GhCCoAOMT7 沉默后，棉花茎秆区域木质部区域颜色明显变浅，木质素含量显著降低，表明GhCCoAOMT7 是木质素合成的关键基因。

Phosphatidylinositol 4 phosphate 5-kinase (PIP5K) is crucial for the phosphatidylinositol (PI) signaling pathway. It plays a significant role in plant growth and development, as well as stress response. However, its effects on cotton are unknown. This study identified PIP5K genes from four cotton species and conducted bioinformatic analyses, with a particular emphasis on the functions of GhPIP5K9a in primary roots. The results showed that cotton PIP5Ks were classified into four subgroups. Analysis of gene structure and motif composition showed obvious conservation within each subgroup. Synteny analysis suggested that the PIP5K gene family experienced significant expansion due to both whole-genome duplication (WGD) and segmental duplication. Transcriptomic data analysis revealed that the majority of GhPIP5K genes had the either low or undetectable levels of expression. Moreover, GhPIP5K9a is highly expressed in the root and was located in plasmalemma. Suppression of GhPIP5K9a transcripts resulted in longer primary roots, longer primary root cells and increased auxin polar transport-related genes expression, and decreased abscisic acid (ABA) content, indicating that GhPIP5K9a negatively regulates cotton primary root growth. This study lays the foundation for further exploration of the role of the PIP5K genes in cotton.

酒棉25号于2023年通过甘肃省农作物品种审定委员会审定,为常规陆地棉品种.根据2020-2021年甘肃省棉花新品种区域试验结果,其生育期139 d,株型紧凑、筒形,株高70.4 cm,第一果枝节位5.4,单株结铃7.5个,铃卵圆形,铃重6.0 g,衣分43.1％,籽指9.5 g,霜前花率93.9％;2年平均666.7 m2籽棉产量、皮棉产量分别为389.9 kg、171.3 kg,比对照酒棉10号分别增产4.6％、15.7％;抗枯萎病,高抗黄萎病;纤维上半部平均长度29.3 mm,断裂比强度28 cN·tex-1,马克隆值4.6,断裂伸长率6.7％,反射率83.9％,黄色深度7.6,长度整齐度指数84.9％,纺纱均匀性指数141.7.介绍了酒棉25号的选育过程、特征特性和栽培技术要点.

The sugar transporter protein (STP) family has been shown to play important roles in plant growth, development, and stress response. However, it has not been studied in cotton compared to other major crops. In this study, we identified 90 STP genes from four cotton species, performed bioinformatic analysis, and focused on the role of GhSTP18 in salt stress. According to our results, cotton STP proteins were divided into four subgroups according to the phylogenetic tree. A synteny analysis suggested that whole-genome duplication (WGD) and segmental duplication were key drivers in the expansion of the STP gene family. The transcriptomic data analysis showed that 29 GhSTP genes exhibited sink-specific expression. Quantitative real time-polymerase chain reaction (qRT-PCR) analyses revealed that expression of GhSTP18 was induced by salt treatment, heat treatment, cold treatment, and drought treatment, and continuously increased during a salt stress time course. Notably, GhSTP18 encodes a plasma membrane-localized galactose transporter. Suppression of GhSTP18 transcription by a virus-induced gene silencing (VIGS) assay reduced sensitivity to salt stress in cotton, indicating that GhSTP18 negatively regulates plant salt tolerance. These results provide an important reference and resource for further studying and deploying STP genes for cotton improvement.

中棉EB004由中国农业科学院棉花研究所选育,在2019-2020年河南省夏棉区域试验和黄河流域棉区早熟常规品种区域试验中综合性状表现为早熟、稳产,吐絮畅,易收摘,高抗枯萎病,耐黄萎病.于2022年通过国家农作物品种审定委员会、河南省主要农作物品种审定委员会审定.介绍了中棉EB004选育过程、特征特性及其关键栽培技术.

由中国农业科学院棉花研究所选育的中棉所141于2021年12月通过国家农作物品种审定委员会审定.该品种早熟性好,丰产性突出,抗枯萎病,耐黄萎病.在介绍中棉所141选育过程、特征特性的基础上,总结了其关键栽培技术.

The AP1/FUL transcription factors are important for floral development, but the underlying molecular mechanisms remain unclear. In this study, we cloned and identified two AP1/FUL-like genes, GhAP1.1 and GhFUL2, in upland cotton, which is a commonly cultivated economically valuable crop. Sequence alignment and phylogenetic analysis indicated that GhAP1.1 and GhFUL2, which are encoded by genes in the AP1/FUL clade, have conserved N-terminal regions but diverse C-terminal domains. Quantitative real-time PCR analysis revealed that GhAP1.1 and GhFUL2 were expressed in the flower and root, and showed opposite expression patterns during shoot apical meristem development. The upregulated expression of GhAP1.1 in Arabidopsis did not result in significant changes to the flowering time or floral organ development, and the transcript levels of the florigen FT increased and those of LFY decreased. Overexpression of GhFUL2 in Arabidopsis delayed flowering and promoted bolting by decreasing FT and LFY transcript levels. Silencing GhFUL2 in cotton dramatically increased the expression of GhFT and GhAP1.3 and promoted flowering. Yeast two-hybrid and bimolecular fluorescence complementation assays indicated that GhAP1.1 could interact with the SVP homolog GhSVP2.2, whereas GhFUL2 formed heterodimers with GhSEP3/GhSEP4 homologs and GhSVP2.2. The present results demonstrated that the functional divergence of GhAP1.1 and GhFUL2, which involved changes in sequences and expression patterns, influenced the regulation of cotton flower development.

Block of proliferation 1 (BOP1) is a key protein that helps in the maturation of ribosomes and promotes the progression of the cell cycle. However, its role in the leaf morphogenesis of cotton remains unknown. Herein, we report and study the function of GhBOP1 isolated from Gossypium hirsutum. The sequence alignment revealed that BOP1 protein was highly conserved among different species. The yeast two-hybrid experiments, bimolecular fluorescence complementation, and luciferase complementation techniques revealed that GhBOP1 interact with GhPES and GhWDR12. Subcellular localization experiments revealed that GhBOP1, GhPES and GhWDR12 were localized at the nucleolus. Suppression of GhBOP1 transcripts resulted in the uneven bending of leaf margins and the presence of young wrinkled leaves by virus-induced gene silencing assay. Abnormal palisade arrangements and the presence of large upper epidermal cells were observed in the paraffin sections of the wrinkled leaves. Meanwhile, a jasmonic acid-related gene, GhOPR3, expression was increased. In addition, a negative effect was exerted on the cell cycle and the downregulation of the auxin-related genes was also observed. These results suggest that GhBOP1 plays a critical role in the development of wrinkled cotton leaves, and the process is potentially modulated through phytohormone signaling.

The natural environment of plants comprises a complex set of biotic and abiotic stresses, and plant responses to these stresses are complex as well. Plant proteomics approaches have significantly revealed dynamic changes in plant proteome responses to stress and developmental processes. Thus, we reviewed the recent advances in cotton proteomics research under changing environmental conditions, considering the progress and challenging factors. Finally, we highlight how single-cell proteomics is revolutionizing plant research at the proteomics level. We envision that future cotton proteomics research at the single-cell level will provide a more complete understanding of cotton's response to stresses.

酒棉23号在甘肃省植棉区种植生育期为135 d,植株筒形,株高73.4 cm,Ⅱ式果枝,单株结铃7.9个,铃重5.7 g,衣分43.7％,籽指11.0 g,霜前花率92.8％,耐枯萎病,高抗黄萎病;2019―2020年甘肃省棉花新品种区域试验中,酒棉23号平均每666.7 m2籽棉产量为359.3 kg,每666.7 m2皮棉产量为160.8 kg.介绍了酒棉23号的选育过程、特征特性、产量、纤维品质、抗病性及栽培技术要点.