Evaluation of drought-tolerant varieties based on root system architecture in cotton (Gossypium hirsutum L.).

BACKGROUND: Root system architecture (RSA) exhibits significant genetic variability and is closely associated with drought tolerance. However, the evaluation of drought-tolerant cotton cultivars based on RSA in the field conditions is still underexplored. RESULTS: So, this study conducted a comprehensive analysis of drought tolerance based on physiological and morphological traits (i.e., aboveground and RSA, and yield) within a rain-out shelter, with two water treatments: well-watered (75 ± 5% soil relative water content) and drought stress (50 ± 5% soil relative water content). The results showed that principal component analysis identified six principal components, including highlighting the importance of root traits and canopy parameters in influencing drought tolerance. Moreover, the systematic cluster analysis was used to classify 80 cultivars into 5 categories, including drought-tolerant cultivars, relatively drought-tolerant cultivars, intermediate cultivars, relatively drought-sensitive cultivars, and drought-sensitive cultivars. Further validation of the drought tolerance index showed that the yield drought tolerance index and biomass drought tolerance index of the drought-tolerant cultivars were 8.97 and 5.05 times higher than those of the drought-sensitive cultivars, respectively. CONCLUSIONS: The RSA of drought-tolerant cultivars was characterised by a significant increase in average length-all lateral roots, a significant decrease in average lateral root emergence angle and a moderate root/shoot ratio. In contrast, the drought-sensitive cultivars showed a significant decrease in average length-all lateral roots and a significant increase in both average lateral root emergence angle and root/shoot ratio. It is therefore more comprehensive and accurate to assess field crop drought tolerance by considering root performance.

Melatonin mitigates cadmium toxicity by promoting root development, delaying root senescence, and regulating cadmium transport in cotton.

Cd ions are absorbed and transported from the soil by crop roots, which are the first organ to be exposed to Cd. This results in an increase in cadmium ions in crops, significantly affecting crop growth and yield. Exogenous melatonin (MT) can help reduce cadmium (Cd) stress in cotton, but the specific contribution of roots to this process remains unclear. In order to address this knowledge gap, an in-situ root phenotyping study was conducted to investigate the the phenotype and lifespan of roots under cadmium stress (Cd) and melatonin treatment (Cd + MT). The results showed that MT alleviated the decreases in plant height, leaf area, SPAD value, stem diameter, stomatal conductance and net photosynthetic rate under Cd stress, which further promoted the biomass accumulation in various cotton organs. What is more, the Cd + MT treatment increased root volume, surface area, and length under Cd stress by 25.63 %, 10.58 %, and 21.89 %, respectively, compared with Cd treatment. Interestingly, compared to Cd treatment, Cd + MT treatment also significantly extended the lifespan of roots and root hairs by 6.68 days and 2.18 days, respectively. In addition, Cd + MT treatment reduced the transport of Cd from roots to shoots, particularly to bolls, and decreased the Cd bioconcentration factor in bolls by 61.17 %, compared to Cd treatment. In conclusion, these findings show that applying MT externally helps reduce Cd stress by delaying root senescence, promoting root development and regulating Cd transport. This method can be an effective approach to managing Cd stress in cotton.

Morphological and Physiological Mechanisms of Melatonin on Delaying Drought-Induced Leaf Senescence in Cotton.

Leaf senescence reduces the photosynthetic capacity of leaves, thus significantly affecting the growth, development, and yield formation of cotton. Melatonin (MT) is a multipotent substance proven to delay leaf senescence. However, its potential mechanism in delaying leaf senescence induced by abiotic stress remains unclear. This study aimed to explore the effect of MT on delaying drought-induced leaf senescence in cotton seedlings and to clarify its morphological and physiological mechanisms. Drought stress upregulated the leaf senescence marker genes, destroyed the photosystem, and led to excessive accumulation of reactive oxygen species (ROS, e.g., H2O2 and O2-), thus accelerating leaf senescence. However, leaf senescence was significantly delayed when 100 µM MT was sprayed on the leaves of the cotton seedlings. The delay was embodied by the increased chlorophyll content, photosynthetic capacity, and antioxidant enzyme activities, as well as decreased H2O2, O2-, and abscisic acid (ABA) contents by 34.44%, 37.68%, and 29.32%, respectively. MT significantly down-regulated chlorophyll degradation-related genes and senescence marker genes (GhNAC12 and GhWRKY27/71). In addition, MT reduced the chloroplast damage caused by drought-induced leaf senescence and maintained the integrity of the chloroplast lamellae structure under drought stress. The findings of this study collectively suggest that MT can effectively enhance the antioxidant enzyme system, improve photosynthetic efficiency, reduce chlorophyll degradation and ROS accumulation, and inhibit ABA synthesis, thereby delaying drought-induced leaf senescence in cotton.

The crucial role of lateral root angle in enhancing drought resilience in cotton.

Introduction: Plant responses to drought stress are influenced by various factors, including the lateral root angle (LRA), stomatal regulation, canopy temperature, transpiration rate and yield. However, there is a lack of research that quantifies their interactions, especially among different cotton varieties. Methods: This experiment included two water treatments: well-watered (75 ± 5% soil relative water content) and drought stress (50 ± 5% soil relative water content) starting from the three-leaf growth stage. Results: The results revealed that different LRA varieties show genetic variation under drought stress. Among them, varieties with smaller root angles show greater drought tolerance. Varieties with smaller LRAs had significantly increased stomatal opening by 15% to 43%, transpiration rate by 61.24% and 62.00%, aboveground biomass by 54% to 64%, and increased seed cotton yield by 76% to 79%, and decreased canopy temperature by 9% to 12% under drought stress compared to the larger LRAs. Varieties with smaller LRAs had less yield loss under drought stress, which may be due to enhanced access to deeper soil water, compensating for heightened stomatal opening and elevated transpiration rates. The increase in transpiration rate promotes heat dissipation from leaves, thereby reducing leaf temperature and protecting leaves from damage. Discussion: Demonstrating the advantages conferred by the development of a smaller LRA under drought stress conditions holds value in enhancing cotton's resilience and promoting its sustainable adaptation to abiotic stressors.

Long-term nitrogen fertilizer management for enhancing use efficiency and sustainable cotton (Gossypium hirsutum L.).

Optimal management of nitrogen fertilizer profoundly impacts sustainable development by influencing nitrogen use efficiency (NUE) and seed cotton yield. However, the effect of long-term gradient nitrogen application on the sandy loam soil is unclear. Therefore, we conducted an 8-year field study (2014-2021) using six nitrogen levels: 0 kg/hm2 (N0), 75 kg/hm2 (N1), 150 kg/hm2 (N2), 225 kg/hm2 (N3), 300 kg/hm2 (N4), and 375 kg/hm2 (N5). The experiment showed that 1) Although nitrogen application had insignificantly affected basic soil fertility, the soil total nitrogen (STN) content had decreased by 5.71%-19.67%, 6.67%-16.98%, and 13.64%-21.74% at 0-cm-20-cm, 20-cm-40-cm, and 40-cm-60-cm soil layers, respectively. 2) The reproductive organs of N3 plants showed the highest nitrogen accumulation and dry matter accumulation in both years. Increasing the nitrogen application rate gradually decreased the dry matter allocation ratio to the reproductive organs. 3) The boll number per unit area of N3 was the largest among all treatments in both years. On sandy loam, the most optional nitrogen rate was 190 kg/hm2-270 kg/hm2 for high seed cotton yield with minimal nitrogen loss and reduced soil environment pollution.

Response of in situ root phenotypes to potassium stress in cotton.

Potassium plays a significant role in the basic functions of plant growth and development. Potassium uptake is closely associated with morphological characteristics of the roots. However, the dynamic characteristics of phenotype and lifespan of cotton (Gossypium hirsutum L.) lateral roots and root hairs under low and high potassium stress remain unclear. In this study, potassium stress experiments (low and high potassium, medium potassium as control) were conducted using RhizoPot (an in situ root observation device) to determine the response characteristics of lateral roots and root hairs in cotton under potassium stress. The plant morphology, photosynthetic characteristics, root phenotypic changes, and lifespan of lateral roots and root hairs were measured. Potassium accumulation, aboveground phenotype, photosynthetic capacity, root length density, root dry weight, root diameter, lateral root lifespan, and root hair lifespan under low potassium stress were significantly decreased compared to medium potassium treatment. However, the root hair length of the former was significantly increased than that of the latter. Potassium accumulation and the lateral root lifespan were significantly increased under high potassium treatment, while root length density, root dry weight, root diameter, root hair length, and root hair lifespan were significantly decreased compared to the medium potassium treatment. Notably, there were no significant differences in aboveground morphology and photosynthetic characters. Principal component analysis revealed that lateral root lifespan, root hair lifespan of the first lateral root, and root hair length significantly correlated with potassium accumulation. The root had similar regularity responses to low and high potassium stress except for lifespan and root hair length. The findings of this study enhance the understanding of the phenotype and lifespan of cotton's lateral roots and root hairs under low and high potassium stress.

Recent advances in methods for in situ root phenotyping.

Roots assist plants in absorbing water and nutrients from soil. Thus, they are vital to the survival of nearly all land plants, considering that plants cannot move to seek optimal environmental conditions. Crop species with optimal root system are essential for future food security and key to improving agricultural productivity and sustainability. Root systems can be improved and bred to acquire soil resources efficiently and effectively. This can also reduce adverse environmental impacts by decreasing the need for fertilization and fresh water. Therefore, there is a need to improve and breed crop cultivars with favorable root system. However, the lack of high-throughput root phenotyping tools for characterizing root traits in situ is a barrier to breeding for root system improvement. In recent years, many breakthroughs in the measurement and analysis of roots in a root system have been made. Here, we describe the major advances in root image acquisition and analysis technologies and summarize the advantages and disadvantages of each method. Furthermore, we look forward to the future development direction and trend of root phenotyping methods. This review aims to aid researchers in choosing a more appropriate method for improving the root system.

Tuning the Defects of Two-Dimensional Layered Carbon/TiO2 Superlattice Composite for a Fast Lithium-Ion Storage.

Defect engineering is one of the effective ways to improve the electrochemical property of electrode materials for lithium-ion batteries (LIB). Herein, an organic functional molecule of p-phenylenediamine is embedded into two-dimensional (2D) layered TiO2 as the electrode for LIB. Then, the 2D carbon/TiO2 composites with the tuning defects are prepared by precise control of the polymerization and carbothermal atmospheres. Low valence titanium in metal oxide and nitrogen-doped carbon nanosheets can be obtained in the carbon/TiO2 composite under a carbonization treatment atmosphere of N2/H2 gas, which can not only increase the electronic conductivity of the material but also provide sufficient electrochemical active sites, thus producing an excellent rate capability and long-term cycle stability. The prepared composite can provide a high capacity of 396.0 mAh g-1 at a current density of 0.1 A g-1 with a high capacitive capacity ratio. Moreover, a high specific capacity of 80.0 mAh g-1 with retention rate of 85% remains after 10,000 cycles at 3.0 A g-1 as well as the Coulomb efficiency close to 100%. The good rate-capability and cycle-sustainability of the layered materials are ascribed to the increase of conductivity, the lithium-ion transport channel, and interfacial capacitance due to the multi-defect sites in the layered composite.

RhizoPot platform: A high-throughput in situ root phenotyping platform with integrated hardware and software.

Quantitative analysis of root development is becoming a preferred option in assessing the function of hidden underground roots, especially in studying resistance to abiotic stresses. It can be enhanced by acquiring non-destructive phenotypic information on roots, such as rhizotrons. However, it is challenging to develop high-throughput phenotyping equipment for acquiring and analyzing in situ root images of root development. In this study, the RhizoPot platform, a high-throughput in situ root phenotyping platform integrating plant culture, automatic in situ root image acquisition, and image segmentation, was proposed for quantitative analysis of root development. Plants (1-5) were grown in each RhizoPot, and the growth time depended on the type of plant and the experimental requirements. For example, the growth time of cotton was about 110 days. The imaging control software (RhizoAuto) could automatically and non-destructively image the roots of RhizoPot-cultured plants based on the set time and resolution (50-4800 dpi) and obtain high-resolution (>1200 dpi) images in batches. The improved DeepLabv3+ tool was used for batch processing of root images. The roots were automatically segmented and extracted from the background for analysis of information on radical features using conventional root software (WinRhizo and RhizoVision Explorer). Root morphology, root growth rate, and lifespan analysis were conducted using in situ root images and segmented images. The platform illustrated the dynamic response characteristics of root phenotypes in cotton. In conclusion, the RhizoPot platform has the characteristics of low cost, high-efficiency, and high-throughput, and thus it can effectively monitor the development of plant roots and realize the quantitative analysis of root phenotypes in situ.

Predicting F v /F m and evaluating cotton drought tolerance using hyperspectral and 1D-CNN.

The chlorophyll fluorescence parameter Fv/Fm is significant in abiotic plant stress. Current acquisition methods must deal with the dark adaptation of plants, which cannot achieve rapid, real-time, and high-throughput measurements. However, increased inputs on different genotypes based on hyperspectral model recognition verified its capabilities of handling large and variable samples. Fv/Fm is a drought tolerance index reflecting the best drought tolerant cotton genotype. Therefore, Fv/Fm hyperspectral prediction of different cotton varieties, and drought tolerance evaluation, are worth exploring. In this study, 80 cotton varieties were studied. The hyperspectral cotton data were obtained during the flowering, boll setting, and boll opening stages under normal and drought stress conditions. Next, One-dimensional convolutional neural networks (1D-CNN), Categorical Boosting (CatBoost), Light Gradient Boosting Machines (LightBGM), eXtreme Gradient Boosting (XGBoost), Decision Trees (DT), Random Forests (RF), Gradient elevation decision trees (GBDT), Adaptive Boosting (AdaBoost), Extra Trees (ET), and K-Nearest Neighbors (KNN) were modeled with F v /F m. The Savitzky-Golay + 1D-CNN model had the best robustness and accuracy (RMSE = 0.016, MAE = 0.009, MAPE = 0.011). In addition, the F v /F m prediction drought tolerance coefficient and the manually measured drought tolerance coefficient were similar. Therefore, cotton varieties with different drought tolerance degrees can be monitored using hyperspectral full band technology to establish a 1D-CNN model. This technique is non-destructive, fast and accurate in assessing the drought status of cotton, which promotes smart-scale agriculture.

Seed Priming With Melatonin Promotes Seed Germination and Seedling Growth of Triticale hexaploide L. Under PEG-6000 Induced Drought Stress.

Melatonin (N-acetyl-5-methoxytryptamine, MT) can mitigate abotic stress, including drought stress on a number of crops. However, it is unclear whether and how seed priming with melatonin alleviates the effects of drought stress on seed germination and seedling growth of triticale (Triticale hexaploide L.). In this study, we investigated the effects of seed priming with MT on seed germination, protective enzyme activity, superoxide anion, and hydrogen peroxide in triticale under PEG-6000 induced drought stress. Seed priming with 20 µM MT alleviated the adverse effects of PEG-6000 induced drought stress on seed germination and seedling growth. Triticale seeds primed with 20 µM MT exhibited improved germination potential, germination rate, germ and radicle length. Specifically, MT priming increased the germination rate by 57.67% compared with unprimed seeds. Seed priming with melatonin also alleviated the adverse effects of PEG-6000 induced drought stress on triticale seedlings. MT pretreatment with 20 µM significantly increased the net photosynthetic rate, transpiration rate, stomatal conductance, plant height, leaf area, and relative chlorophyll concentration, enhanced the activities of superoxide dismutase and peroxidase, and decreased reactive oxygen species (ROS) and malonaldehyde content in the seeds (germ and radicle) and seedlings (leaf and root). Collectively, these results suggest that seed priming with melatonin promotes ROS scavenging capacity and enhances energy supply and antioxidant enzyme activities to alleviate the adverse effects of drought stress in triticale.

In situ Root Phenotypes of Cotton Seedlings Under Phosphorus Stress Revealed Through RhizoPot.

Phosphorus (P) deficiency is a common challenge in crop production because of its poor mobility through the soil. The root system plays a significant role in P absorption from the soil and is the initial indicator of low P levels. However, the phenotypic dynamics and longevity of cotton roots under P stress remain unknown. In this study, RhizoPot, an improvised in situ root observation device, was used to monitor the dynamics of root phenotypes of cotton seedlings under P-deficient (PD) and P-replete (PR) conditions. Low P stress reduced P absorption and accumulation in the roots, leading to low dry weight accumulation. Cotton seedlings responded to low P stress by increasing the number of lateral roots, specific root length, branch density, root length density, and length of root hairs. Additionally, the life span of root hairs was prolonged. Low P stress also reduced the average diameter of roots, promoted root extension, expanded the root coverage area, and increased the range of P acquisition. Principal component analysis revealed that the net root growth rate, root length density, root dry weight, P absorption efficiency, average root hair length, and taproot daily growth significantly influenced the cotton root architecture. Collectively, these results show that low P stress reduces the net growth rate of cotton seedling roots and restricts plant growth. Plants respond to P deficiency by extending the life span of root hairs and increasing specific root length and lateral root branch density. This change in root system architecture improves the adaptability of plants to low P conditions. The findings of this study may guide the selection of cotton varieties with efficient P utilization.

Physiological and Comparative Transcriptomic Analysis Provide Insight Into Cotton (Gossypium hirsutum L.) Root Senescence in Response.

Nitrogen (N) deficiency is one of the pivotal environmental factors that induce leaf senescence. However, little is known regarding the impact of low N on root senescence in cotton. Thus, the objective of this study was to investigate the effect of low nitrogen on root senescence. In this study, the molecular mechanism of cotton root senescence in response to nitrogen deficiency was investigated by combing physiological and transcriptomic analysis when no nitrogen and normal nitrogen (138mg N·kg-1 soil). The results showed that: (1) nitrogen starvation induced the premature senescence of leaf, while delaying root senescence. (2) The increase in catalase (CAT) activity at 60, 80, and 100days after emergence (DAE), combined with decrease of malonaldehyde content at 60, 80, and 100 DAE, and the content of abscisic acid (ABA), all of these contributed to the delay of root senescence by low nitrogen treatment. (3) To study the molecular mechanisms underlying root senescence, the gene expression profiling between low nitrogen and normal nitrogen treatments were compared pairwise at 20, 40, 60, 80, and 100 DAE. A total of 14,607 genes were identified to be differentially expressed at these five points. (5) Most genes involved in glutathione (GSH) and ascorbate peroxidase (APX) synthesis were upregulated, while ABA, apoptosis, caspase, and cell cycle-related differentially expressed genes (DEGs) were downregulated. Coupled with the physiology data, these results provide new insights into the effect of nitrogen starvation on root senescence.