Cotton is an important economic crop in China, and the current cotton industry is transitioning toward mechanization, with chemical defoliation being a key factor in this development. Breeding defoliant-sensitive cultivars can promote rapid and concentrated leaf abscission, effectively reduce trash content in seed cotton, improve mechanical harvesting efficiency, and advance the mechanization process of cotton production. In this study, we employed bulked segregant analysis (BSA) and transcriptome profiling to elucidate the molecular mechanisms underlying cotton defoliation. BSA-seq analysis identified nine major loci associated with defoliation. Integrating RNA-seq and tissue-specific expression profiles, we screened 22 differentially expressed genes (DEGs) that are highly expressed in stem and leaf organs. Based on expression pattern analysis, haplotype analysis, studies on the regulatory relationship between the gene and ethylene, and functional annotation of Arabidopsis homologs, we selected the cotton homolog GhAMT1;2, located at ChrD11: 56738327-56740667 bp, as the core candidate gene. This gene encodes a root high-affinity ammonium transporter involved in nitrogen metabolism. Subsequent virus-induced gene silencing (VIGS) experiments demonstrated that GhAMT1;2 exhibits downregulated expression following defoliant treatment and positively regulates cotton defoliation. This study successfully mapped defoliation-associated loci and validated gene function, providing a theoretical foundation for breeding and improving machine-harvestable cotton varieties.

Chemical defoliation is essential for mechanized harvesting of cotton (Gossypium hirsutum L.), yet the molecular mechanisms governing abscission zone (AZ) cell fate and cell-wall remodeling remain unresolved. Here, single-nucleus RNA sequencing (snRNA-seq) coupled with Monocle2 trajectory analysis delineates a pseudotemporal transition of protective-layer cells from C16_0 to C16_1, which coincides with AZ maturation. We identify the cell-wall glycoprotein GhSKS6 as a central hub in this differentiation process. CRISPR/Cas9-mediated knockout of GhSKS6 impairs AZ fracture-layer formation, delays leaf abscission, and downregulates genes associated with hemicellulose and xyloglucan remodeling. Subcellular localization and plasmolysis assays confirm the residency of GhSKS6 in the cell wall, while its overexpression in protoplasts accelerates cell wall regeneration. Moreover, the NAC transcription factor GhNAC47 directly binds to the GhSKS6 promoter to activate its expression, thereby modulating cotton leaf abscission. Our findings reveal a regulatory module involving GhNAC47 and GhSKS6 that coordinates protective-layer formation and cell-wall remodeling, offering precise molecular targets for breeding machine-harvested cotton cultivars.

The calcineurin B-like (CBL)-interacting protein kinase (CIPK) signaling network is the core regulatory node in the response to abiotic stress in plants; it regulates plant homeostasis by regulating various proteins mediating ion transport. However, there are few reports on CIPK-mediated ion transporters in the cotton (Gossypium hirsutum) response to drought stress. Through yeast two-hybrid assays, we identified SUPPRESSOR OF K+ TRANSPORT GROWTH DEFECT 1 (GhSKD1), which interacts with GhCIPK6D1. GhSKD1 was significantly up-regulated after drought stress, while GhSKD1 localized to the cell membrane and nucleus. Functional studies revealed that GhSKD1 positively regulates K+ efflux, thus enhancing drought tolerance in cotton. Genetic and biochemical evidence showed that the phosphorylation of GhSKD1 by GhCIPK6D1 mediates K+ influx in guard cells, thereby regulating stomatal aperture and drought tolerance in cotton. GhSKD1 represents a previously uncharacterized protein that mediates potassium ion transport during the drought stress response. This finding identifies another target of CIPK regulation in the CPL-CIPK signaling network and provides insights into the mechanisms of drought tolerance in plants.

Chemical defoliation stands as the ultimate tool in enabling the mechanical harvest of cotton, offering economic and environmental advantages. However, the underlying molecular mechanism that triggers leaf abscission through defoliant remains unsolved. In this study, we meticulously constructed a transcriptomic atlas through single-nucleus mRNA sequencing (snRNA-seq) of the abscission zone (AZ) from cotton petiole. We identified two newly-formed cell types, abscission cells and protection layer cells in cotton petiole AZ after defoliant treatment. GhRLF1 (RAPID LEAF FALLING 1), as one of the members of the cytokinin oxidase/dehydrogenase (CKX) gene family, was further characterized as a key marker gene unique to the abscission cells following defoliant treatment. Overexpression of GhRLF1 resulted in reduced cytokinin accumulation and accelerated leaf abscission. Conversely, CRISPR/Cas9-mediated loss of GhRLF1 function appeared to delay this process. Its interacting regulators, GhWRKY70, acting as "Pioneer" activator, and GhMYB108, acting as "Successor" activator, orchestrate a sequential modulation of GhWRKY70/GhMYB108-GhRLF1-CTK (cytokinin) within the AZ to regulate cotton leaf abscission. GhRLF1 not only regulates leaf abscission but also reduces cotton yield. Consequently, transgenic lines that exhibit rapid leaf falling and require less defoliant but show unaffected cotton yield were developed for mechanical harvesting. This was achieved using a defoliant-induced petiole-specific promoter, proPER21, to drive GhRLF1 (proPER21::RLF1). This pioneering biotechnology offers a new strategy for the chemical defoliation of machine-harvested cotton, ensuring stable production and reducing leaf debris in harvested cotton, thereby enhancing environmental sustainability.

Global water scarcity and extreme weather intensify drought stress, significantly reducing cotton yield and quality worldwide. Drought treatments are conducted using a population of chromosome segment substitution lines generated from E22 (G. hirsutum) and 3-79 (G. barbadense) as parental lines either show superior yields or fiber quality under both control and drought conditions. Fourteen datasets, covering 4 yields and 4 quality traits, are compiled and assessed for drought resistance using the drought resistance coefficient (DRC) and membership function value of drought resistance (MFVD). Genome-wide association studies, linkage analysis, and bulked segregant analysis are combined to analyze the DR-related QTL. A total of 121 significant QTL are identified by DRC and MFVD of the 8 traits. CRISPR/Cas9 and virus-induced gene silencing techniques verified DRR1 and DRT1 as pivotal genes in regulating drought resistant of cotton, with hap3-79 exhibiting greater drought resistance than hapE22 concerning DRR1 and DRT1. Moreover, 14 markers with superior yield and fiber quality are selected for drought treatment. This study offers valuable insights into yield and fiber quality variations between G. hirsutum and G. barbadense amid drought, providing crucial theoretical and technological backing for developing cotton varieties resilient to drought, with high yield and superior fiber quality.

The occurrence of whole-genome duplication or polyploidy may promote plant adaptability to harsh environments. Here, we clarify the evolutionary relationship of eight GhCIPK6 homologous genes in upland cotton (Gossypium hirsutum). Gene expression and interaction analyses indicate that GhCIPK6 homologous genes show significant functional changes after polyploidy. Among these, GhCIPK6D1 and GhCIPK6D3 are significantly up-regulated by drought stress. Functional studies reveal that high GhCIPK6D1 expression promotes cotton drought sensitivity, while GhCIPK6D3 expression promotes drought tolerance, indicating clear functional differentiation. Genetic and biochemical analyses confirm the synergistic negative and positive regulation of cotton drought resistance through GhCBL1A1-GhCIPK6D1 and GhCBL2A1-GhCIPK6D3, respectively, to regulate stomatal movement by controlling the directional flow of K+ in guard cells. These results reveal differentiated roles of GhCIPK6 homologous genes in response to drought stress in upland cotton following polyploidy. The work provides a different perspective for exploring the functionalization and subfunctionalization of duplicated genes in response to polyploidization.

Edible oils with high unsaturated fatty acids, particularly oleic acid, are beneficial to human health. Cotton is one of the top five oil crops in the world, but the mechanism of high-quality oil synthesis and regulatory networks in cotton are largely unclear. Here, we identified Leafy cotyledon1-like 1 (GhL1L1), a NF-YB subfamily gene that is specifically expressed during somatic embryogenesis and seed maturation in cotton. Overexpression of GhL1L1 regulates the contents of unsaturated fatty acids in cotton, especially in the seeds, which is associated with altered expression of the cotton fatty acid biosynthesis-related genes. GhL1L1 synergistically enhanced the expression of GhFAD2-1A by binding to the G-box in its promoter, leading to an increase in the content of linoleic acid. Furthermore, this activation could be enhanced by GhNF-YC2 and GhNF-YA1 by form a transcriptional complex. Collectively, these results contribute to provide new insights into the molecular mechanism of oil biosynthesis in cotton and can facilitate genetic manipulation of cotton varieties with enhanced oil content.

SummaryN6‐methyladenosine (m6A) is the most prevalent internal modification present in mRNAs, and is considered to participate in a range of developmental and biological processes. Drought response is highly regulated at the genomic, transcriptional and post‐transcriptional levels. However, the biological function and regulatory mechanism of m6A modification in the drought stress response is still poorly understood. We generated a transcriptome‐wide m6A map using drought‐resistant and drought‐sensitive varieties of cotton under different water deficient conditions to uncover patterns of m6A methylation in cotton response to drought stress. The results reveal that m6A represents a common modification and exhibit dramatic changes in distribution during drought stress. More 5'UTR m6A was deposited in the drought‐resistant variety and was associated with a positive effect on drought resistance by regulating mRNA abundance. Interestingly, we observed that increased m6A abundance was associated with increased mRNA abundance under drought, contributing to drought resistance, and vice versa. The demethylase GhALKBH10B was found to decrease m6A levels, facilitating the mRNA decay of ABA signal‐related genes (GhZEP, GhNCED4 and GhPP2CA) and Ca2+ signal‐related genes (GhECA1, GhCNGC4, GhANN1 and GhCML13), and mutation of GhALKBH10B enhanced drought resistance at seedling stage in cotton. Virus‐induced gene silencing (VIGS) of two Ca2+‐related genes, GhECA1 and GhCNGC4, reduced drought resistance with the decreased m6A enrichment on silenced genes in cotton. Collectively, we reveal a novel mechanism of post‐transcriptional modification involved in affecting drought response in cotton, by mediating m6A methylation on targeted transcripts in the ABA and Ca2+ signalling transduction pathways.

Plant anthers are composed of different specialized cell types with distinct roles in plant reproduction. High temperature (HT) stress causes male sterility, resulting in crop yield reduction. However, the spatial expression atlas and regulatory dynamics during anther development and in response to HT remain largely unknown. Here, the first single-cell transcriptome atlas and chromatin accessibility survey in cotton anther are established, depicting the specific expression and epigenetic landscape of each type of cell in anthers. The reconstruction of meiotic cells, tapetal cells, and middle layer cell developmental trajectories not only identifies novel expressed genes, but also elucidates the precise degradation period of middle layer and reveals a rapid function transition of tapetal cells during the tetrad stage. By applying HT, heterogeneity in HT response is shown among cells of anthers, with tapetal cells responsible for pollen wall synthesis are most sensitive to HT. Specifically, HT shuts down the chromatin accessibility of genes specifically expressed in the tapetal cells responsible for pollen wall synthesis, such as QUARTET 3 (QRT3) and CYTOCHROME P450 703A2 (CYP703A2), resulting in a silent expression of these genes, ultimately leading to abnormal pollen wall and male sterility. Collectively, this study provides substantial information on anthers and provides clues for heat-tolerant crop creation.

Root and leaf are essential organs of plants in sensing and responding to drought stress. However, comparative knowledge of non-coding RNAs (ncRNAs) of root and leaf tissues in the regulation of drought response in cotton is limited. Here, we used deep sequencing data of leaf and root tissues of drought-resistant and drought-sensitive cotton varieties for identifying miRNAs, lncRNAs and circRNAs. A total of 1531 differentially expressed (DE) ncRNAs was identified, including 77 DE miRNAs, 1393 DE lncRNAs and 61 DE circRNAs. The tissue-specific and variety-specific competing endogenous RNA (ceRNA) networks of DE lncRNA-miRNA-mRNA response to drought were constructed. Furthermore, the novel drought-responsive lncRNA 1 (DRL1), specifically and differentially expressed in root, was verified to positively affect phenotypes of cotton seedlings under drought stress, competitively binding to miR477b with GhNAC1 and GhSCL3. In addition, we also constructed another ceRNA network consisting of 18 DE circRNAs, 26 DE miRNAs and 368 DE mRNAs. Fourteen circRNA were characterized, and a novel molecular regulatory system of circ125- miR7484b/miR7450b was proposed under drought stress. Our findings revealed the specificity of ncRNA expression in tissue- and variety-specific patterns involved in the response to drought stress, and uncovered novel regulatory pathways and potentially effective molecules in genetic improvement for crop drought resistance.

INTRODUCTION:Drought is the principal abiotic stress that severely impacts cotton (Gossypium hirsutum) growth and productivity. Upon sensing drought, plants activate stress-related signal transduction pathways, including ABA signal and mitogen-activated protein kinase (MAPK) cascade. However, as the key components with the fewest members in the MAPK cascade, the function and regulation of GhMKKs need to be elucidated. In addition, the relationship between MAPK module and the ABA core signaling pathway remains incompletely understood. OBJECTIVE:Here we aim to elucidate the molecular mechanism of cotton response to drought, with a focus on mitogen-activated protein kinase (MAPK) cascades activating ABA signaling. METHODS:Biochemical, molecular and genetic analysis were used to study the GhMAP3K62-GhMKK16-GhMPK32-GhEDT1 pathway genes. RESULTS:A nucleus- and membrane-localized MAPK cascade pathway GhMAP3K62-GhMKK16-GhMPK32, which targets and phosphorylates the nuclear-localized transcription factor GhEDT1, to activate downstream GhNCED3 to mediate ABA-induced stomatal closure and drought response was characterized in cotton. Overexpression of GhMKK16 promotes ABA accumulation, and enhances drought tolerance via regulating stomatal closure under drought stress. Conversely, RNAi-mediated knockdown of GhMKK16 expression inhibits ABA accumulation, and reduces drought tolerance. Virus-induced gene silencing (VIGS)-mediated knockdown of either GhMAP3K62, GhMPK32 or GhEDT1 expression represses ABA accumulation and reduces drought tolerance through inhibiting stomatal closure. Expression knockdown of GhMPK32 or GhEDT1 in GhMKK16-overexpressing cotton reinstates ABA content and stomatal opening-dependent drought sensitivity to wild type levels. GhEDT1 could bind to the HD boxes in the promoter of GhNCED3 to activate its expression, resulting in ABA accumulation. We propose that the MAPK cascade GhMAP3K62-GhMKK16-GhMPK32 pathway functions on drought response through ABA-dependent stomatal movement in cotton.

以果园典型路面不平度的采集为研究对象,建立基于激光雷达点云处理的路面不平度采集方法,搭建了基于三维激光雷达的路面不平度采集系统平台,结合点云处理技术完成路面高程信息的提取;采用AR(au?to regressive,自回归)模型依据比例分析法对路面功率谱密度进行计算,确定不平度等级,并通过加速度振动记录仪进行系统验证,利用系统开展典型果园路面不平度数据信息采集试验.试验结果显示,果园路面不平度结果为水泥路面主要集中在B级,B级占比82.33％;砂石路面主要集中在C级,C级占比84.00％;泥土路面主要集中在D、E等级,D级路面占比48.67％,E级占比31.00％,表明基于三维激光雷达采集系统与数据处理方法在果园路面不平度采集和评价上是可行的.最终不平度评价结果显示,三维激光雷达果园路面不平度采集系统应用可靠,评价结果准确,适合于山地林、果、茶园路面不平度的采集.

In order to understand the molecular mechanism of cotton's response to drought during the flowering and boll stage, transcriptomics and metabolomics were carried out for two introgression lines (drought-tolerant line: T307; drought-sensitive line: S48) which were screened from Gossypium hirsutum cv. ‘Emian22’ with some gene fragments imported from Gossypium barbadense acc. 3–79, under drought stress by withdrawing water at flowering and boll stage. Results showed that the basic drought response in cotton included a series of broad-spectrum responses, such as amino acid synthesis, hormone (abscisic acid, ABA) signal transduction, and mitogen-activated protein kinases signal transduction pathway, which activated in both drought-tolerant and drought-sensitive lines. However, the difference of their imported fragments and diminished sequences triggers endoplasmic reticulum (ER) protein processing, photosynthetic-related pathways (in leaves), and membrane solute transport (in roots) in drought-tolerant line T307, while these are missed or not activated in drought-sensitive line S48, reflecting the different drought tolerance of the two genotypes. Virus-induced gene silencing assay of drought-tolerant differentially expressed heat shock protein (HSP) genes (mainly in leaf) and ATP-binding cassette (ABC) transporter genes (mainly in roots) indicated that those genes play important role in cotton drought tolerant. Combined analysis of transcriptomics and metabolomics highlighted the important roles of ER-stress-related HSP genes and root-specific ABC transporter genes in plants drought tolerance. These results provide new insights into the molecular mechanisms underlying the drought stress adaptation in cotton.

植物根际微生物群落对植物生长和适逆性至关重要,本研究对干旱条件下棉花根际真菌群落进行分析,旨在探明干旱胁迫对棉花根际真菌多样性和群落结构的影响,为利用有益微生物提高棉花水分利用率提供理论依据。以陆地棉Jin 668 （Gossypium hirsutum cv. Jin668）为试验材料,采用盆栽控水方式,对处于开花期的棉花根际土壤（SDP）和未种植棉花土壤（SOPD）进行干旱处理,正常浇水的棉花根际土壤（SPN）和无棉花土壤（SNPN）为对照。从中采集土壤样品,提取DNA,采用IlluminaMiseq对真菌ITS1区域进行高通量测序,研究土壤中真菌多样性。结果共鉴定到970个OTUs,SNPN、SOPD、SPN和SDP样品中真菌OTUs数量分别为481、528、743和752个,其中288个OTUs为所有组共有。对获得OTUs进行门、纲、目、科和属5个分类水平的划分表明,棉花根际真菌群落结构主要由子囊菌门（82.70%）和担子菌门（10.15%）组成;干旱处理使粪壳菌纲（Sordariomycetes）、粪壳菌目（Sordariales）和毛壳菌科（Chaetomiaceae）丰度显著降低,而散囊菌目（Eurotiales）、发菌科（Trichocomaceae）、曲霉属（Aspergillus）和青霉属（Penicillum）的丰度显著增加。多样性分析结果显示,与未种棉花的土壤相比,有棉花的土壤中真菌群落的α多样性显著增加;同时, SPN和SDP之间的真菌群落结构更相似,而与SNPN和SOPD间差异较大。研究表明,棉花根际存在丰富的真菌群落,干旱对土壤中真菌的丰度和多样性有显著影响。本研究从微生物的角度为提高棉花耐旱性的研究提供新见解。

为实现苗圃田间果树钵苗的快速搬运,研制了一种适用于苗圃田间的自走式电动双轨运输机.针对运输机的应用环境、果树钵苗质量大小等需求,分析提出运输机应具备的装载量、电机功率、运输速度等设计参数;并根据技术参数要求,提出运输机的总体结构并对关键部件进行设计;最后,对运输机进行了实验室与田间试验.试验结果显示,运输机载重能力为750 kg,空载每百米能耗为9.16 W·h,满载每百米能耗为27.6 W·h,运行速度为0.564～0.718 m/s,运输机从起步到匀速行驶所用时间小于2.5 s,紧急制动距离小于0.05 m,振动加速度振幅总体小于0.4 g.以上结果表明,研制的运输机可以满足实现苗圃田间作业需求,可以有效提高作业效率、降低劳动强度.

为解决山地果园运输机在实际使用中因路况起伏不平而引起的剧烈振动问题,结合山地实际路面情况,设计使用CDC阻尼器的半主动悬架系统,并安装于华南农业大学研发的丘陵山地果园电动轮式运输机.以安装使用CDC阻尼器的半主动悬架系统的丘陵山地果园轮式运输机为研究对象,采用振动仪和振动传感器搭建振动测试系统,在行驶速度和载荷一定的工况下,分别测试该轮式运输机装有半主动悬架前后行驶过程中座椅位置Z轴的振动信号,考察半主动悬架装车前后的振动差异.结果显示,装有使用CDC阻尼器的半主动悬架的轮式运输机振动降幅达50％,达到了半主动悬架系统的设计要求;装有使用CDC阻尼器的半主动悬架系统的车身振动频率集中,范围大于8 Hz,表明车辆的驾驶舒适性较好.

Wettability of graphene is important for researching, modulating the wettability at the same time as producing deserve more attention. In this work, a fast, convenient and low-cost preparation method was introduced to produce superhydrophobic graphene with a biomimetics surface structure that was inspired from taro leaves. First, the surface structure of the taro leaf was obtained by the scanning electron microscopy (SEM). Then a regular structure imitating a taro leaf was designed. Subsequently, a graphene layer with biomimetic surface structure similar to the taro leaf was induced on the surface of polyimide film by Nd:YAG laser. The morphology of the graphene surface was adjusted by the scanning speed and laser fluence during laser processing under the contribution of the photothermal effect. Under the optimal processing parameters for wettability, wetting tests with the water, sweat and serum droplets showed that the graphene with biomimetic surface has superhydrophobicity, anti-sweat and serum adhesion properties with the same contact angle and sliding angle of 153° and 3.3°, respectively. This optimized biomimetics laser-induced graphene can be used to protect exposed graphene-based devices from liquid erosion.

The mitogen-activated protein kinase (MAPK) cascade pathway, which has three components, MAP3Ks, MKKs and MPKs, is involved in diverse biological processes in plants. In the current study, MAPK cascade genes were identified in three cotton species, based on gene homology with Arabidopsis. Selection pressure analysis of MAPK cascade genes revealed that purifying selection occurred among the cotton species. Expression pattern analysis showed that some MAPK cascade genes differentially expressed under abiotic stresses and phytohormones treatments, and especially under drought stress. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) experiments showed extensive interactions between different MAPK cascade proteins. Virus-induced gene silencing (VIGS) assays showed that some MAPK cascade modules play important roles in the drought stress response, and the GhMAP3K14–GhMKK11–GhMPK31 signal pathway was demonstrated to regulate drought stress tolerance in cotton. This study provides new information on the function of MAPK cascade genes in the drought response, and will help direct molecular breeding for improved drought stress tolerance in cotton. Following a comprehensive analysis of MAPK cascade genes in cotton, we show that the GhMAP3K14–GhMKK11–GhMPK31 pathway is involved in the response to drought stress.

针对目前山地林果茶园挖穴作业人工劳动强度大和作业效率低的问题,设计了一种具有行驶动力的山地林果茶园电动自走式挖穴机.该挖穴机能自动完成钻头进给行程与复位行程.田间试验结果表明,该挖穴机单次作业平均挖穴时间为50.7 s,单次有效挖穴作业能耗为6.38 W·h,施肥穴平均深度为392.5 mm,施肥穴平均直径为303 mm,行走速度为1.237 m/s,能够顺利通过15°斜坡.挖穴作业过程中钻头的进给与回程运动均无需人工操作,可提高工作效率,降低劳动强度,并保证挖穴操作中的安全性.