Biomolecular condensates, such as stress and germ granules, often contain subcompartments. For instance, the Caenorhabditis elegans germ granule, which localizes near the outer nuclear membrane of germ cell nuclei, is composed of at least four ordered compartments, each housing distinct sets of proteins and RNAs. How these compartments form and why they are spatially ordered remains poorly understood. Here, we show that the conserved DEAD-box RNA helicase DDX-19 defines another compartment of the larger C. elegans germ granule, which we term the D compartment. The D compartment exhibits properties of a liquid condensate and forms between the outer nuclear pore filament and other compartments of the germ granule. Two nuclear pore proteins, NPP-14 and GLEL-1, are required for its formation, suggesting that the D compartment localizes adjacent to the outer nuclear membrane through interactions with the nuclear pore. The loss of DDX-19, NPP-14 or GLEL-1 leads to functional defects, including aberrant formation of the other four germ granule compartments, a loss of germline immortality and dysregulation of small RNA-based transgenerational epigenetic inheritance programs. Hence, we propose that a function of the D compartment is to anchor larger germ granules to nuclear pores, enabling germ granule compartmentalization and promoting transgenerational RNA surveillance.

Heartwood are mainly used as Castanopsis hystrix valuable wood, which play a vital role in determining the quality of wood for sales, value assessment, and consumption. There are two defects in the heartwood produced by C. hystrix commercial forests: a low proportion of heartwood, reducing the output of high-value wood, and the uneven color of the heartwood from red to reddish-brown, affecting its visual appearance and decorative performance. As these two defects seriously influence the quality of C. hystrix heartwood, breeding of high-yield and high-quality heartwood varieties is critical. The present study examined 20-year-old C. hystrix forests in Yulin, Liuzhou, and Pingxiang counties in China. Through analysis of the proportion, density, brightness L* , chroma a* , and chroma b* of heartwood, genetic variation in each trait was described, genetic parameters were estimated, and the genotype by environment interaction effect was clarified. The average value of the above traits were 34.76%, 0.67 g/cm(3), 52.22, 13.90, and 17.25, respectively. The results of the joint three-site variance analysis reveal that heartwood proportion, density, chroma a* , chroma b* have significant (p < 0.05) or extremely significant (p < 0.01) genotype, environment, and their interaction effects, with family heritability values of 0.25, 0.23, 0.31, and 0.54, respectively. Based on the BLUP-GGE biplot of single and multiple traits, the families with strong adaptability and stable genetics were selected. The average genetic gains in heartwood proportion, density, chroma a* , and chroma b* from families 7, 8 and 9 were 5.83%, 0.02%, 1.04%, and 3.37%, respectively. The three families selected in this study were vital for promotion and utilization, providing a theoretical foundation for the innovation of C. hystrix germplasm and precise improvement of wood quality in South China.

Biomolecular condensates, such as the nucleolus, stress granules/processing bodies and germ granules, are multiphase assemblages whose formation mechanisms and significance remain poorly understood. Here we identify protein constituents of the spatiotemporally ordered P, Z and M multiphase condensates in Caenorhabditis elegans germ granules using optimized TurboID-mediated proximity biotin labelling. These include 462, 41 and 86 proteins localizing to P, Z and M condensates, respectively, of which 522 were previously unknown protein constituents. Each condensate’s proteins are enriched for distinct classes of structured and intrinsically disordered domains, suggesting divergent functions and assembly mechanisms. Through a functional screen, we identify a germ granule protein, HERD-1, which prevents the mixing of P, Z and M condensates. Mixing in herd-1 mutants correlates with disorganization of germline small RNA pathways and prolonged epigenetic inheritance of RNA interference-induced gene silencing. Forced mixing of these condensate components using a nanobody with specific binding activity against green fluorescent protein also extends epigenetic inheritance. We propose that active maintenance of germ granule immiscibility helps to organize and regulate small RNA-driven transgenerational epigenetic inheritance in C. elegans. Zhao et al. report that the Z compartment protein HERD-1 regulates transgenerational epigenetic inheritance in Caenorhabditis elegans at least in part by preventing the mixing of germ granule condensates.

Biomolecular condensates, such as nucleolus, stress granules/P bodies, and germ granules in Caenorhabditis elegans , are multiphase assemblages. However, the mechanisms underlying the formation of these multi-phase condensates and their significance remain poorly understood. Here, we demonstrate that the spatially ordered PZMS assemblage of germ granules in Caenorhabditis elegans is established hierarchically by three LOTUS domain proteins. Additionally, we identified a Z granules protein termed HERD-1 that prevents the mixing of P granules, Z granules, and Mutator foci. Surprisingly, loss of herd-1 leads to RNAi defect but paradoxically results in prolonged transgenerational epigenetic inheritance of RNAi for over 50 generations due to uncontrolled pUG RNAs-siRNAs cycles after PZM fusion. Moreover, ectopic recruitment of Mutator foci to P granules or Z granules closely mimics the phenotype observed in herd-1 mutant. Our findings propose an assembly principle for multi-phase condensates and highlight the active maintenance of immiscibility between condensates by specific cellular machinery, which restrict transgenerational epigenetic inheritance.

Cold stress is a potent abiotic stress resulting in cotton yield and fiber quality loss. We hypothesized that GhGTG1 could increase low temperature stress tolerance by improving the sensitivity to ABA. Genome-wide identification, biochemical traits and functional characterization of GhGTG1, were investigated under different levels of ABA and cold stress. GhGTG1 had two highly conserved functional domains (ABA_GPCR, GPHR_N) in all cotton species. Under ABA treatment, over expression of GhGTG1 in Arabidopsis had a lower germination rate and root elongation. While under 4 degrees C stress, transgenic plants had a higher germination rate and root elongation. This indicate that GhGTG1 improved sensitivity to ABA under cold stress. In transgenic plants, CAT and SOD activity was increased, while H2O2 and MDA content decreased. Less trypan blue and DAB staining area suggested that GhGTG1 enhances cold stress resistance. Under 4 degrees C treatment, silence cotton seedlings through VIGS method, resulted in a poor growth, lower relative leaf water content, chlorophyll content but a higher ion leakage and excised leaf water loss. The activity of CAT and SOD decreased, H2O2 and MDA content was increased. Two marker genes (OST1 and ABCG40) are associated with the stomatal movement, but these genes were substantially suppressed. This suggesting that GhGTG1 can induce cold stress tolerance by improving ABA sensitivity Via., maintaining stomatal movement. These data will provide new insight of cold resistance mechanism in cotton and further will help the breeder to develop new cotton cold stress tolerance germplasm.

Background: Cotton is mainly grown for its natural fiber and edible oil. The fiber obtained from cotton is the indispensable raw material for the textile industries. The ever changing climatic condition, threatens cotton production due to a lack of sufficient water for its cultivation. Effects of drought stress are estimated to affect more than 50% of the cotton growing regions. To elucidate the drought tolerance phenomenon in cotton, a backcross population was developed from G. tomentosum, a drought tolerant donor parent and G. hirsutum which is highly susceptible to drought stress.Results: A genetic map of 10 888 SNP markers was developed from 200 BC 2 F 2 populations. The map spanned 4 191.3 centi-Morgan（c M）, with an average distance of 0.104 7 c M, covering 51% and 49% of At and Dt sub genomes, respectively. Thirty stable Quantitative trait loci（QTLs） were detected, in which more than a half were detected in the At subgenome. Eighty-nine candidate genes were mined within the QTL regions for three traits:cell membrane stability（CMS）, saturated leaf weight（SLW） and chlorophyll content. The genes had varied physiochemical properties. A majority of the genes were interrupted by introns, and only 15 genes were intronless,accounting for 17% of the mined genes. The genes were found to be involved molecular function（MF）, cellular component（CC） and biological process（BP）, which are the main gene ontological（GO） functions. A number of mi RNAs were detected, such as mi R164, which is associated with NAC and MYB genes, with a profound role in enhancing drought tolerance in plants. Through RT-q PCR analysis, 5 genes were found to be the key genes involved in enhancing drought tolerance in cotton. Wild cotton harbors a number of favorable alleles, which can be exploited to aid in improving the narrow genetic base of the elite cotton cultivars. The detection of 30 stable QTLs and 89 candidate genes found to be contributed by the donor parent, G. tomentosum, showed the significant genes harbored by the wild progenitors which can be exploited in developing more robust cotton genotypes with diverse tolerance levels to various environmental stresses.Conclusion: This was the first study involving genome wide association mapping for drought tolerance traits in semi wild cotton genotypes. It offers an opportunity for future exploration of these genes in developing highly tolerant cotton cultivars to boost cotton production.

In this investigation, whole-genome identification and functional characterization of the cotton dehydrin genes was carried out. A total of 16, 7, and 7 dehydrin proteins were identified in G. hirsutum, G. arboreum and G. raimondii, respectively. Through RNA sequence data and RT-qPCR validation, Gh_A05G1554 (GhDHN_03) and Gh_D05G1729 (GhDHN_04) were highly upregulated, and knockdown of the two genes, significantly reduced the ability of the plants to tolerate the effects of osmotic and salt stress. The VIGS-plants recorded significantly higher concentration levels of oxidants, hydrogen peroxide (H2O2) and malondialdehyde (MDA), furthermore, the four stress responsive genes GhLEA2, Gh_D12G2017 (CDKF4), Gh_A07G0747 (GPCR) and a transcription factor, trihelix, Gh_A05G2067, were significantly downregulated in VIGS-plants, but upregulated in wild types under osmotic and salt stress condition. The result indicated that dehydrin proteins are vital for plants and can be exploited in developing a more osmotic and salt stress-resilient germplasm to boost and improve cotton production.

Segregation distortion (SD) is a phenomenon common among stable or segregating populations, and the principle behind it is still an issue that puzzles many researchers. An F 2:3 generations developed from the wild cotton species of the D genomes was applied to investigate the possible plant transcription factors within the segregation distortion regions (SDRs). We constructed a consensus map between two maps in the D genome, map A; Gossypium klotzschianum and Gossypium davidsonii and Map B; Gossypium thurberi and Gossypium trilobum . The two maps were developed from 188 F 2:3 populations for each map, a total of 1492 markers, were linked to the 13 linkage groups. The consensus linkage map size was 1467.445 cM with an average marker distance of 1.0370cM. Chr02 had the highest percentage of segregation distortion with 58.621% followed by Chr07 with 47.887%. A total of 6,038 genes were mined within the segregation distortion regions (SDR region) of Chr02 and Chr07 with 2,308 gene in Chr02 and 3,730 genes in Chr07, we obtained a total of 1,117 domains within the SDR with a total of 622 domains shared between the two chromosomes, the first 9 domains all belonged to the plant resistance genes (R genes), the largest domain was PF00069 with a total of 188 genes. A total of 287 miRNAs were found to target the various genes, such as gr-miR398, gra-miR5207, miR164a, miR164b, miR164c among others which have been found to target top-ranked stress-responsive transcription factors such as NAC genes. Moreover, the genes were found to be regulated by various stress responsive cis -regulatory elements. RNA profiling showed that higher numbers of genes were highly upregulated in abiotic and different fiber development stages. The result shows that the SDR regions could be playing an important role in the evolution of significant genes in plants.

[目的]对棉花D基因组中Bet v 1基因家族进行分析,比较其在不同抗性棉种间的表达模式差异,为深入研究Bet v 1基因在棉花抗黄萎病中的作用提供理论依据.[方法]通过生物信息学方法对D基因组中Bet v 1基因进行鉴定.通过雷蒙德氏棉(Gossypium raimondii,D5)、三裂棉(G.trilobum,D8)和瑟伯氏棉(G.thurberi,D1)转录组和实时荧光定量聚合酶链式反应(qRT-PCR)分析Bet v 1基因在黄萎病菌处理下的表达模式.利用病毒诱导的基因沉默技术(VIGS)对Bet v 1基因进行功能鉴定.[结果][棉花D基因组中包含59个成员,其中57个基因带有内含子,分布于8条染色体上,多数为亲水性蛋白并定位于细胞质.]黄萎病菌胁迫条件下3个野生棉种的Bet v 1基因表达量与其抗病水平一致.将不同表达水平的基因分为3组,其中第3组基因响应黄萎病菌侵染并在抗病棉种瑟伯氏棉中高表达,表明该组基因可能与黄萎病胁迫应答反应有关.从中筛选出高水平表达的Bet v 1基因,在陆地棉中沉默相应的同源基因,棉株感病加重,揭示该基因在棉花抵御黄萎病菌侵染过程中起正调控作用.[结论]响应黄萎病菌胁迫的Bet v 1基因在棉花抗黄萎病复杂的生物过程中至关重要.本研究结果为棉花Bet v 1家族基因的深入研究提供依据,为进一步解析棉花Bet v 1基因的功能奠定基础.

We found 33, 17, and 20 Alba genes in Gossypium hirsutum, Gossypium arboretum, and Gossypium raimondii, respectively. The Alba protein lengths ranged from 62 to 312 aa, the molecular weight (MW) from 7.003 to 34.55 kDa, grand average hydropathy values of -1.012 to 0.609 and isoelectric (pI) values of -3 to 11. Moreover, miRNAs such as gra-miR8770 targeted four genes, gra-miR8752 and gra-miR8666 targeted three genes, and each and gra-miR8657 a, b, c, d, e targeted 10 genes each, while the rests targeted 1 to 2 genes each. Similarly, various cis-regulatory elements were detected with significant roles in enhancing abiotic stress tolerance, such as CBFHV (RYCGAC) with a role in cold stress acclimation among others. Two genes, Gh_D01G0884 and Gh_D01G0922, were found to be highly induced under water deficit and salt stress conditions. Through virus-induced gene silencing (VIGS), the VIGS cotton plants were found to be highly susceptible to both water deficit and salt stresses; the VIGS plants exhibited a significant reduction in root growth, low cell membrane stability (CMS), saturated leaf weight (SLW), chlorophyll content levels, and higher excised leaf water loss (ELWL). Furthermore, the stress-responsive genes and ROS scavenging enzymes were significantly reduced in the VIGS plants compared to either the wild type (WT) and or the positively controlled plants. The VIGS plants registered higher concentration levels of hydrogen peroxide and malondialdehyde, with significantly lower levels of the various antioxidants evaluated an indication that the VIGS plants were highly affected by salt and drought stresses. This result provides a key foundation for future exploration of the Alba proteins in relation to abiotic stress.

We identified 672, 374, and 379 CYPs proteins encoded by the CYPs genes in Gossypium hirsutum, Gossypium raimondii, and Gossypium arboreum, respectively. The genes were found to be distributed in all 26 chromosomes of the tetraploid cotton, with chrA05, chrA12, and their homeolog chromosomes harboring the highest number of genes. The physiochemical properties of the proteins encoded by the CYP450 genes varied in terms of their protein lengths, molecular weight, isoelectric points (pI), and even grand hydropathy values (GRAVY). However, over 99% of the cotton proteins had GRAVY values below 0, which indicated that the majority of the proteins encoded by the CYP450 genes were hydrophilic in nature, a common property of proteins encoded by stress-responsive genes. Moreover, through the RNA interference (RNAi) technique, the expression levels of Gh_D07G1197 and Gh_A13G2057 were suppressed, and the silenced plants showed a higher concentration of hydrogen peroxide (H2O2) with a significant reduction in the concentration levels of glutathione (GSH), ascorbate peroxidase (APX), and proline compared to the wild types under drought and salt stress conditions. Furthermore, the stress-responsive genes 1-Pyrroline–5-Carboxylate Synthetase (GhP5CS), superoxide dismutase (GhSOD), and myeloblastosis (GhMYB) were downregulated in VIGS plants, but showed upregulation in the leaf tissues of the wild types under drought and salt stress conditions. In addition, CYP450-silenced cotton plants exhibited a high level of oxidative injury due to high levels of oxidant enzymes, in addition to negative effects on CMS, ELWL, RLWC, and chlorophyll content The results provide the basic foundation for future exploration of the proteins encoded by the CYP450 genes in order to understand the physiological and biochemical mechanisms in enhancing drought and salt stress tolerance in plants.

Verticillium wilt that is caused by Verticillium dahliae, does result in massive annual yield losses and fiber quality decline in cotton. Control by conventional mechanisms is not possible due to a wide host range and the longevity of dormant fungi in the soil in the case of absence of a suitable host. Plants have developed various mechanisms to boost their immunity against various diseases, and one is through the induction of various genes. In this research, we carried out RNA sequencing and then identified the members of the adenosine triphosphate (ATP)-binding cassette (ABC) proteins to be critical in enhancing resistance to V. dahliae infection. A total of 166 proteins that are encoded by the ABC genes were identified in Gossypium raimondii with varying physiochemical properties. A novel ABC gene, Gorai.007G244600 (ABCF5), was found to be highly upregulated, and its homolog in the tetraploid cotton Gh_D11G3432 (ABCF5), was then silenced through virus induced gene silencing (VIGS) in G. hirsutum, tetraploid upland cotton. The mutant cotton seedlings ability to tolerate V. dahliae infection was significantly reduced. Based on the evaluation of oxidant enzymes, hydrogen peroxide (H2O2) and malondialdehyde (MDA) showed significantly increased levels in the leaves of the mutant compared to the wild type. In addition, antioxidant enzymes, peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD) concentrations were reduced in the mutant cotton leaves after treatment with V. dahliae fungi as compared to the wild type. Moreover, expression levels of the biotic stress genes, cotton polyamine oxidase (GhPAO), cotton ribosomal protein L18 (GhRPL18), and cotton polygalacturonase-inhibiting protein-1 (GhPGIP1), were all downregulated in the mutant but they were highly upregulated in the various tissues of the wild cotton seedlings. This research has shown that ABC genes could play an important role in enhancing the immunity of cotton to V. dahliae infection, and thus can be explored in developing more resilient cotton genotypes with improved resistance to V. dahliae infection in cotton.

BACKGROUND:Auxins play an important role in plant growth and development; the auxins responsive gene; auxin/indole-3-acetic acid (Aux/IAA), small auxin-up RNAs (SAUR) and Gretchen Hagen3 (GH3) control their mechanisms. The GH3 genes function in homeostasis by the catalytic activities in auxin conjugation and bounding free indole-3-acetic acid (IAA) to amino acids.RESULTS:In our study, we identified the GH3 genes in three cotton species; Gossypium hirsutum, Gossypium arboreum and Gossypium raimondii, analyzed their chromosomal distribution, phylogenetic relationships, cis-regulatory element function and performed virus induced gene silencing of the novel Gh_A08G1120 (GH3.5) gene. The phylogenetic tree showed four clusters of genes with clade 1, 3 and 4 having mainly members of the GH3 of the cotton species while clade 2 was mainly members belonging to Arabidopsis. There were no paralogous genes, and few orthologous genes were observed between Gossypium and other species. All the GO terms were detected, but only 14 genes were found to have described GO terms in upland cotton, more biological functions were detected, as compared to the other functions. The GH3.17 subfamily harbored the highest number of the cis-regulatory elements, most having promoters towards dehydration-responsiveness. The RNA expression analysis revealed that 10 and 8 genes in drought and salinity stress conditions respectively were upregulated in G. hirsutum. All the genes that were upregulated in plants under salt stress conditions were also upregulated in drought stress; moreover, Gh_A08G1120 (GH3.5) exhibited a significant upregulation across the two stress factors. Functional characterization of Gh_A08G1120 (GH3.5) through virus-induced gene silencing (VIGS) revealed that the VIGS plants ability to tolerate drought and salt stresses was significantly reduced compared to the wild types. The chlorophyll content, relative leaf water content (RLWC), and superoxide dismutase (SOD) concentration level were reduced significantly while malondialdehyde concentration and ion leakage as a measure of cell membrane stability (CMS) increased in VIGS plants under drought and salt stress conditions.CONCLUSION:This study revealed the significance of the GH3 genes in enabling the plant's adaptation to drought and salt stress conditions as evidenced by the VIGS results and RT-qPCR analysis.

Drought, salinity and cold stresses have a major impact on cotton production, thus identification and utilization of plant genes vital for plant improvement Whole-genome identification and functional characterizations of the IQ67-domain (IQD) protein family was carried out in which 148, 77, and 79 IQD genes were identified in Gossypium hirsutum, G. raimondii, and G. arboreum. The entire IQD proteins had varied physiochemical properties, however; their grand hydropathy values were negative, which demonstrated that the proteins were hydrophilic, a property common among the proteins encoded by various stresses responsive genes, such as the late embryogenesis abundant (LEA) proteins. The IQD proteins were predicted to be majorly sublocalized in the nucleus; moreover, various cis-regulatory elements with higher role in enhancing abiotic stress tolerance were detected. RNA-seq and RT-qPCR analysis revealed two key genes, Gh_D06G0014 and Gh_A09G1608 with significantly higher upregulation across the various tissues under drought, salt and cold stress. Knockdown of the two genes negatively affected the ability of G. hirsutum to tolerate the effects of the three stress factors, being all the antioxidant assayed were significantly low concentrations compared to the oxidizing enzymes in VIGS plants under stress, furthermore, morphological and physiological traits were all negatively affected in VIGS plants. Expression levels of GhLEA2, GhCDK_F4, GPCR (TOM1) and Gh_A05G2067 (TH), the stress responsive genes were all downregulated in the VIGS plants, but significantly upregulated in WT and positively controlled plants. The results demonstrated that the IQD genes could be responsible for enhancing drought, salt and cold stress tolerance in cotton.

Background The efficient detection and initiation of appropriate response to abiotic stresses are important to plants survival. The plant G-protein coupled receptors (GPCRs) are diverse membranous proteins that are responsible for signal transduction. Results In this research work, we identified a novel gene of the GPCR domain, transformed and carried out the functional analysis in Arabidopsis under drought and cold stresses. The transgenic lines exposed to drought and cold stress conditions showed higher germination rate, increased root length and higher fresh biomass accumulation. Besides, the levels of antioxidant enzymes, glutathione (GSH) and ascorbate peroxidase (APX) exhibited continuously increasing trends, with approximately threefold higher than the control, implying that these ROS-scavenging enzymes were responsible for the detoxification of ROS induced by drought and cold stresses. Similarly, the transgenic lines exhibited stable cell membrane stability (CMS), reduced water loss rate in the detached leaves and significant values for the saturated leaves compared to the wild types. Highly stress-responsive miRNAs were found to be targeted by the novel gene and based on GO analysis; the protein encoded by the gene was responsible for maintaining an integral component of membrane. In cotton, the virus-induced gene silencing (VIGS) plants exhibited a higher susceptibility to drought and cold stresses compared to the wild types. Conclusion The novel GPCR gene enhanced drought and cold stress tolerance in transgenic Arabidopsis plants by promoting root growth and induction of ROS scavenging enzymes. The outcome showed that the gene had a role in enhancing drought and cold stress tolerance, and can be further exploited in breeding for more stress-resilient and tolerant crops.

We identified 102, 51 and 51 proteins encoded by the trihelix genes in Gossypium hirsutum, Gossypium arboreum and Gossypiumraimondii, respectively. RNA sequence data and real-time quantitative polymerase chain reaction analysis showed that Gh_A05G2067 (GT-2) was highly upregulated under drought and salt stress conditions. Transient expression of GT-2-green fluorescent protein fusion protein in protoplast showed that GT-2 was localized in the nucleus. The overexpression of GT-2 conferred an enhanced drought tolerance to cotton, with lower malondialdehyde, hydrogen peroxide contents and higher reactive oxygen scavenging enzyme activities. Moreover, chlorophyll content, relative leaf water content (RLWC), excised leaf water loss (ELWL) and cell membrane stability (CMS) were relatively stable in the GT-2-overexpressed lines compared to wild-type (WT). Similarly, stress-responsive genes RD29A, SOS1, ABF4 and CBL1 were highly upregulated in the GT-2-overexpressed lines but were significantly downregulated in WT. In addition, the GT-2-silenced cotton plants exhibited a high level of oxidation injury, due to high levels of oxidant enzymes, in addition to negative effects on CMS, ELWL, RLWC and chlorophyll content. These results mark the foundation for future exploration of the trihelix genes in cotton, with an aim of developing more resilient, versatile and highly tolerant cotton genotypes.

Abiotic stresses have negative effects on plants growth and development. Plants, being sessile, have developed specific adaptive strategies that allow them to rapidly detect and respond to abiotic stress factors. The detoxification efflux carriers (DTX)/multidrug and toxic compound extrusion (MATE) transporters are of significance in the translocation of abscisic acid (ABA), a phytohormone with profound role in plants under various abiotic stress conditions. The ABA signaling cascades are the core regulators of abiotic stress responses in plants, triggering major changes in gene expression and adaptive physiological responses. We therefore carried out genome-wide analysis of the DTX/MATE gene family, transformed a DTX/MATE gene in Arabidopsis and carried out functional analysis under drought, salt, and cold stress conditions. We identified 128, 70, and 72 DTX/MATE genes in Gossypium hirsutum, Gossypium arboreum, and Gossypium raimondii, respectively. The proteins encoded by the DTX/MATE genes showed varied physiochemical properties but they all were hydrophobic. The Gh_D06G0281 (DTX/MATE) over-expressing Arabidopsis lines were highly tolerant under drought, salt, and cold stress with high production of antioxidant enzymes and significantly reduced levels of oxidants. Lipid peroxidation, as measured by the level of malondialdehyde concentrations was relatively low in transgenic lines compared to wild types, an indication of reduced oxidative stress levels in the transgenic plants. Based on physiological measurements, the transgenic plants exhibited significantly higher relative leaf water content, reduced excised leaf water loss and a significant reduction in ion leakage as a measure of the cell membrane stability compared to the wild types. Abiotic stress responsive genes, ABF4, CBL1, SOS1, and RD29B were highly expressed in the transgenic lines compared to the non-transformed wild type plants. The protein encoded by the Gh_D06G0281 (DTX/MATE) gene was predicted to be located within the plasma membrane. Since signals from extracellular stimuli are transmitted through the plasma membrane most of which are conducted by plasma membrane proteins it is possible the Gh_D06G0281 (DTX/MATE) gene product could be important for this process.

Nitrogen is a key macronutrient needed by plants to boost their production, but the development of cotton genotypes through conventional approaches has hit a bottleneck due to the narrow genetic base of the elite cotton cultivars, due to intensive selection and inbreeding. Based on our previous research, in which the BC2F2 generations developed from two upland cotton genotypes, an abiotic stress-tolerant genotype, G. tomentosum (donor parent) and a highly-susceptible, and a highly-susceptible, but very productive, G. hirsutum (recurrent parent), were profiled under drought stress conditions. The phenotypic and the genotypic data generated through genotyping by sequencing (GBS) were integrated to map drought-tolerant quantitative trait loci (QTLs). Within the stable QTLs region for the various drought tolerance traits, a nodule-inception-like protein (NLP) gene was identified. We performed a phylogenetic analysis of the NLP proteins, mapped their chromosomal positions, intron-exon structures and conducted ds/dn analysis, which showed that most NLP genes underwent negative or purifying selection. Moreover, the functions of one of the highly upregulated genes, Gh_A05G3286 (Gh NLP5), were evaluated using the virus gene silencing (VIGS) mechanism. A total of 226 proteins encoded by the NLP genes were identified, with 105, 61, and 60 in Gossypium hirsutum, G. raimondii, and G. arboreum, respectively. Comprehensive Insilico analysis revealed that the proteins encoded by the NLP genes had varying molecular weights, protein lengths, isoelectric points (pI), and grand hydropathy values (GRAVY). The GRAVY values ranged from a negative one to zero, showing that proteins were hydrophilic. Moreover, various cis-regulatory elements that are the binding sites for stress-associated transcription factors were found in the promoters of various NLP genes. In addition, many miRNAs were predicted to target NLP genes, notably miR167a, miR167b, miR160, and miR167 that were previously shown to target five NAC genes, including NAC1 and CUC1, under N-limited conditions. The real-time quantitative polymerase chain reaction (RT-qPCR) analysis, revealed that five genes, Gh_D02G2018, Gh_A12G0439, Gh_A03G0493, Gh_A03G1178, and Gh_A05G3286 were significantly upregulated and perhaps could be the key NLP genes regulating plant response under N-limited conditions. Furthermore, the knockdown of the Gh_A05G3286 (GhNLP5) gene by virus-induced silencing (VIGS) significantly reduced the ability of these plants to the knockdown of the Gh_A05G3286 (GhNLP5) gene by virus-induced gene silencing (VIGS) significantly reduced the ability of the VIGS-plants to tolerate N-limited conditions compared to the wild types (WT). The VIGS-plants registered lower chlorophyll content, fresh shoot biomass, and fresh root biomass, addition to higher levels of malondialdehyde (MDA) and significantly reduced levels of proline, and superoxide dismutase (SOD) compared to the WT under N-limited conditions. Subsequently, the expression levels of the Nitrogen-stress responsive genes, GhTap46, GhRPL18A, and GhKLU were shown to be significantly downregulated in VIGS-plants compared to their WT under N-limited conditions. The downregulation of the nitrogen-stress responsive genes provided evidence that the silenced gene had an integral role in enhancing cotton plant tolerance to N-limited conditions.

本研究基于13个表型特征对9个国家的208份海岛棉材料进行了遗传多样性分析.试验显示,208份海岛棉表型性状平均变异系数18.33％,平均多样性指数1.30.对不同来源海岛棉表型性状进行多重比较分析发现,伸长率和马克隆值差异不显著,铃形差异显著,其他性状差异极显著.对各性状间的相关分析发现38对性状极显著相关,12对性状显著相关.通过因子分析将表型性状简化为5个互不相关的主成分,累计贡献率达81.69％.通过聚类分析,将208份材料在阈值21.5处划分为6个类群.试验表明:海岛棉种质各性状具有广泛变异性和多样性;中国主产区海岛棉材料遗传多样性最为丰富,但产量性状较差,美洲海岛棉材料的纤维品质优势明显;表型性状的5个主成分分别为纤维品质因子、棉铃特征因子、果枝特征因子、株高因子和马克隆值因子;各类群具有明显的区域性和品质差异性.这些结果为拓宽中国海岛棉的遗传基础,提高中国海岛棉育种效率有重要参考价值.