Identification and Expression Analysis of the NPF Genes in Cotton

The NPF (NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER FAMILY) transports various substrates, including nitrogen (N), which is essential for plant growth and development. Although many NPF homologs have been identified in various plants, limited studies on these proteins have been reported in cotton. This study identified 75, 71, and 150 NPF genes in Gossypium arboreum, G. raimondii, and G. hirsutum, respectively, via genome-wide analyses. The phylogenetic tree indicated that cotton NPF genes are subdivided into eight subgroups, closely clustered with Arabidopsis orthologues. The chromosomal location, gene structure, motif compositions, and cis-elements have been displayed. Moreover, the collinearity analysis showed that whole-genome duplication event has played an important role in the expansion and diversification of the NPF gene family in cotton. According to the transcriptome and qRT-PCR analyses, several GhNPFs were induced by the nitrogen deficiency treatment. Additional functional experiments revealed that virus-induced silencing (VIGS) of the GhNPF6.14 gene affects the growth and N absorption and accumulation in cotton. Thus, this study lays the foundation for further functional characterization of NPF genes in cotton

Ameliorative Effects of Silicon against Salt Stress in Gossypium hirsutum L.

Silicon (Si) could alleviate the adverse effect of salinity in many crops, but the effect in cotton remains unclear. In this study, we evaluated the role of Si in regulating the salt stress tolerance of cotton by analyzing the induced morpho-physiological changes. A hydroponic experiment was conducted by using contrasting salt-tolerant cotton genotypes (sensitive Z0102; tolerant Z9807) and four treatments (CK, control; CKSi, 0.4 mM Si; NaCl, 150 mM NaCl; NaClSi, 150 mM NaCl+0.4 mM Si). The results showed that Si significantly enhanced the net photosynthesis rate and improved the growth of cotton seedling under salt stress in both salt-sensitive and salt-tolerant genotypes. Exogenous Si significantly reduced the accumulation of reactive oxygen species (ROS) and decreased the malondialdehyde (MDA) content in salt-stressed cotton. In addition, the application of Si up-regulated the expression of CAT1, SODCC and POD, and significantly enhanced the antioxidant enzymatic activities, such as catalase (CAT) and peroxidase (POD), of the salt-stressed cotton seedlings. Further, Si addition protected the integrity of the chloroplast ultrastructure, including key enzymes in photosynthesis such as ferredoxin-NADP reeducates (FNR), ATP synthase (Mg2+Ca2+-ATPase) and ribulose-1, 5-bisphosphate carboxylase/oxygenase (RubisCO), and the structure and function of the photosynthetic apparatus PSII from salt stress. Moreover, Si significantly increased the effective stomatal density and stomatal aperture in the salt-stressed cotton seedlings. Taken together, Si could likely ameliorate adverse effects of salt stress on cotton by improving the ROS scavenging ability and photosynthetic capacity

Phosphorus Availability Affects the Photosynthesis and Antioxidant System of Contrasting Low-P-Tolerant Cotton Genotypes

Phosphorus (P) is an essential macronutrient, and an important component of plant metabolism. However, little is known about the effects of low P availability on P absorption, the photosynthetic electron transport chain, and the antioxidant system in cotton. This study used cotton genotypes (sensitive FJA and DLNTDH and tolerant BX014 and LuYuan343) with contrasting low-P tolerance in a hydroponic experiment under 15 µM, 50 µM, and 500 μM P concentrations. The results showed that low P availability reduced plant development and leaf area, shoot length, and dry weight in FJA and DLNADH, compared to BX014 and LuYuan343. The low P availability decreased the gas-exchange parameters such as the net photosynthetic rate, transpiration rate, and stomatal conductance, and increased the intercellular CO2 concentration. Chlorophyll a fluorescence demonstrated that the leaves’ absorption and trapped-energy flux were largely steady. In contrast, considerable gains in absorption and trapped-energy flux per reaction center resulted from decreases in the electron transport per reaction center under low-P conditions. In addition, low P availability reduced the activities of antioxidant enzymes and increased the content of malondialdehyde in the cotton genotypes, especially in FJA and DLNTDH. Moreover, low P availability reduced the activity of PEPC and generated a decline in the content of ATP and NADPH. Our research can provide a theoretical physiological basis for the growth and tolerance of cotton under low-P conditions

Systematic characterization of Gossypium GLN family genes reveals a potential function of GhGLN1.1a regulates nitrogen use efficiency in cotton

Abstract The enzyme glutamine synthetase (GLN) is mainly responsible for the assimilation and reassimilation of nitrogen (N) in higher plants. Although the GLN gene has been identified in various plants, there is little information about the GLN family in cotton (Gossypium spp.). To elucidate the roles of GLN genes in cotton, we systematically investigated and characterized the GLN gene family across four cotton species (G. raimondii, G. arboreum, G. hirsutum, and G. barbadense). Our analysis encompassed analysis of members, gene structure, cis-element, intragenomic duplication, and exploration of collinear relationships. Gene duplication analysis indicated that segmental duplication was the primary driving force for the expansion of the GhGLN gene family. Transcriptomic and quantitative real-time reverse-transcription PCR (qRT-PCR) analyses indicated that the GhGLN1.1a gene is responsive to N induction treatment and several abiotic stresses. The results of virus-induced gene silencing revealed that the accumulation and N use efficiency (NUE) of cotton were affected by the inactivation of GhGLN1.1a. This study comprehensively analyzed the GhGLN genes in Gossypium spp., and provides a new perspective on the functional roles of GhGLN1.1a in regulating NUE in cotton

DataSheet_2_Genetic variation in morphological traits in cotton and their roles in increasing phosphorus-use-efficiency in response to low phosphorus availability.xlsx

Phosphorus (P) is an essential macronutrient required for fundamental processes in plants. Trait plasticity is crucial for plant adaptation to environmental change. Variations in traits underlie diverse phosphorus (P) acquisition strategies among plants. Nevertheless, how the intraspecific plasticity and integration of morphological traits contribute to Phosphorus-Use-Efficiency (PUE) in cotton is unknown. In this study, 25 morphological traits were evaluated in 384 cotton genotypes grown with low P (LP, 10μmol. L−1) and normal nutrition (CK, 500μmol. L−1) to assess the genetic variability of morphological traits and their relationship to phosphorus use efficiency. Results revealed a large genetic variation in mostly morphological traits under low P. Significant enhancement in root traits and phosphorus efficiency-related traits like PUE was observed at LP as compared to CK conditions. In response to low P availability, cotton genotypes showed large plasticity in shoot and total dry biomass, phosphorus, and nitrogen efficiency-related traits (i.e., phosphorus/nitrogen use efficiency, phosphorus/nitrogen uptake efficiency), and most root traits, but a limited response in root dry biomass, taproot length, root surface area, root volume, and SPAD value. In addition, significant correlations were observed between PUtE (phosphorus uptake efficiency), NUE (nitrogen use efficiency), TDB (total dry biomass), and RTD (root tissue density) with PUE under both P supply level and phosphorus stress index, which may be a key indicator for improving PUE under LP conditions. Most root traits are most affected by genotypes than nutrition level. Conserved PUE is more affected by the nutrition level than the genotype effect. Principal component analysis depicted the comprehensive indicators under two P supply conditions were mainly reflected in root-related traits and morphological indicators such as dry matter biomass. These results indicate that interspecific variations exist within these cotton genotypes and traits. Our study provides suggestions for future research to enhance the ability of the earth system model to predict how crops respond to environmental interference and provide target quality for cotton breeding in phosphorus-deficient areas.</p

DataSheet_1_Genetic variation in morphological traits in cotton and their roles in increasing phosphorus-use-efficiency in response to low phosphorus availability.docx

Phosphorus (P) is an essential macronutrient required for fundamental processes in plants. Trait plasticity is crucial for plant adaptation to environmental change. Variations in traits underlie diverse phosphorus (P) acquisition strategies among plants. Nevertheless, how the intraspecific plasticity and integration of morphological traits contribute to Phosphorus-Use-Efficiency (PUE) in cotton is unknown. In this study, 25 morphological traits were evaluated in 384 cotton genotypes grown with low P (LP, 10μmol. L−1) and normal nutrition (CK, 500μmol. L−1) to assess the genetic variability of morphological traits and their relationship to phosphorus use efficiency. Results revealed a large genetic variation in mostly morphological traits under low P. Significant enhancement in root traits and phosphorus efficiency-related traits like PUE was observed at LP as compared to CK conditions. In response to low P availability, cotton genotypes showed large plasticity in shoot and total dry biomass, phosphorus, and nitrogen efficiency-related traits (i.e., phosphorus/nitrogen use efficiency, phosphorus/nitrogen uptake efficiency), and most root traits, but a limited response in root dry biomass, taproot length, root surface area, root volume, and SPAD value. In addition, significant correlations were observed between PUtE (phosphorus uptake efficiency), NUE (nitrogen use efficiency), TDB (total dry biomass), and RTD (root tissue density) with PUE under both P supply level and phosphorus stress index, which may be a key indicator for improving PUE under LP conditions. Most root traits are most affected by genotypes than nutrition level. Conserved PUE is more affected by the nutrition level than the genotype effect. Principal component analysis depicted the comprehensive indicators under two P supply conditions were mainly reflected in root-related traits and morphological indicators such as dry matter biomass. These results indicate that interspecific variations exist within these cotton genotypes and traits. Our study provides suggestions for future research to enhance the ability of the earth system model to predict how crops respond to environmental interference and provide target quality for cotton breeding in phosphorus-deficient areas.</p