Effects of N application methods on cotton yield and fertilizer N recovery efficiency in salinity fields with drip irrigation under mulch film using 15N tracing technique.

Introduction: Drip irrigation under mulch film promotes a non-uniform salinity distribution in salt fields. The effect of different N application methods on the growth and yield of cotton under drip irrigation under mulch film conditions in eastern coastal saline-alkaline soils in China remain remained unclear. Methods: A randomized complete block design was used in the experiment. Three N application methods were assigned: N applied under mulch film (low-salinity area; UM), N applied between mulch films (high-salinity area; BM), and half N applied under mulch film and half between mulch films (HUHB). Results: Plant height, photosynthesis, Chl content, boll load, biomass, boll weight and boll density under UM were all significantly higher than those under the other two treatments. The N absorption of UM was higher than in the other two treatments, which might be attributed to the expression of GHNRT1.5 and GHNRT2.1. The net NO3- influx in the roots in UM increased significantly compared with that in BM. The yield and FNRE of UM were 3.9% and 9.1%, respectively, and were 26.52% and 90.36% higher than under HUHB and BM treatments. Discussion: UM not only improved cotton yield but also alleviated the pollution of N residue on drip irrigation under mulch film conditions in salt areas.

C-terminally encoded peptides act as signals to increase cotton root nitrate uptake under nonuniform salinity.

Soil salinity is often heterogeneous in saline fields. Nonuniform root salinity increases nitrate uptake into cotton (Gossypium hirsutum) root portions exposed to low salinity, which may be regulated by root portions exposed to high salinity through a systemic long-distance signaling mechanism. However, the signals transmitted between shoots and roots and their precise molecular mechanisms for regulating nitrate uptake remain unknown. Here, we showed that nonuniform root salinity treatment using split-root systems increases the expression of C-TERMINALLY ENCODED PEPTIDE (GhCEP) genes in high-saline-treated root portions. GhCEP peptides originating in high-saline-treated root portions act as ascending long-distance mobile signals transported to the shoots to promote the expression of CEP DOWNSTREAM (GhCEPD) genes by inducing the expression of CEP receptor (GhCEPR) genes. The shoot-derived GhCEPD polypeptides act as descending mobile signals transported to the roots through the phloem, increasing the expression of nitrate transport genes NITRATE TRANSPORTER 1.1 (GhNRT1.1), GhNRT2.1, and GhNRT1.5 in nonsaline-treated root portions, thereby increasing nitrate uptake in the nonsaline-treated root portions. This study indicates that GhCEP and GhCEPD signals are transported between roots and shoots to increase nitrate uptake in cotton, and the transport from the nonsaline root side is in response to nonuniform root salinity distribution.

Light induced shoot-sourced transcription factor HY5 regulates the nitrate uptake of cotton by shoot-to-root signal transport.

Elongated hypocotyls 5 (HY5) is a transcription factor that can be induced by illumination and promotes nitrate uptake in Arabidopsis. However, whether GhHY5 regulates nitrate uptake in cotton is unknown. In this study, the cotton seedlings growing in light and dark conditions were treated with 15N-labeled nutrient solution to study whether the GhHY5 regulates nitrate uptake in cotton. The results showed that the 15N content and GhNRT1.1 expression in the light condition were higher than that in the dark condition, indicating that light induced the expression of GhNRT1.1 and subsequently promoted N uptake. Additionally, the expression of GhHY5 in the leaf and root of cotton was induced by light and the expression pattern of GhHY5 in the root was similar to that of GhNRT1.1. Furthermore, when the GhHY5 expression in the root was reduced, the 15N content and GhNRT1.1 expression were both decreased, indicating that the GhNRT1.1 expression was regulated by GhHY5. The root expression of GhHY5 was decreased in the grafted seedlings which the GhHY5 in the shoot was silenced by VIGS or the seedlings which the hypocotyl was girdled, but the expression of GhHY5 on one side root of the grafted cotton seedling was not changed if the GhHY5 was silenced on the other side root. Thus, we proposed that the light induced shoot-derived GhHY5 gene or GhHY5 protein may be transported from the xylem to the root, regulating the expression of GhHY5 and GhNRT1.1, and thus regulating N uptake at the root of cotton.

Seeding depth and seeding rate regulate apical hook formation by inducing GhHLS1 expression via ethylene during cotton emergence.

Apical hook formation is essential for the emergence and stand establishment of cotton plants. Searching for agronomic measures to regulate apical hook formation and clarifying its mechanism are important for full stand establishment in cotton. In this study, cotton seeds were sown at varying seeding rates or depths in sand to determine if and how apical hook formation was regulated by seeding rates or depths. The results showed that deep seeding or low seeding rates increased mechanical pressure and then increased ethylene content by increasing GhACO1 and GhACS2 expression to improve apical hook formation. Silencing of the GhACO1 and GhACS2 genes or exogenous application of 1-methylcyclopropene (1-MCP) decreased the ethylene content and inhibited apical hook formation in the cotton seedlings. Deep seeding, a low seeding rate, or 1-amino cyclopropane-1-carboxylic acid (ACC) treatment increased the expression of GhHLS1 and GhPIF3 genes, but their expression was decreased in theVIGS-ACO1 and VIGS-ACS2 seedlings. Silencing of the GhHLS1 and GhPIF3 genes inhibited apical hook formation, although the expression of GhACO1 and GhACS2 was unchanged. GhPIF3 may act upstream of GhHLS1, as the expression of GhPIF3 in the VIGS-HLS1 seedlings was unchanged, while the expression of GhHLS1 in the VIGS-PIF3 seedlings decreased. These results suggested that raised mechanical pressure could increase ethylene content by inducing GhACO1 and GhACS2 gene expression, which promoted apical hook formation by increasing the expression of GhHLS1. Therefore, adjusting the mechanical pressure through changing the seeding depth or seeding rate is an important means to regulate apical hook formation and emergence.

Global gene expression in cotton (Gossypium hirsutum L.) leaves to waterlogging stress.

Cotton is sensitive to waterlogging stress, which usually results in stunted growth and yield loss. To date, the molecular mechanisms underlying the responses to waterlogging in cotton remain elusive. Cotton was grown in a rain-shelter and subjected to 0 (control)-, 10-, 15- and 20-d waterlogging at flowering stage. The fourth-leaves on the main-stem from the top were sampled and immediately frozen in liquid nitrogen for physiological measurement. Global gene transcription in the leaves of 15-d waterlogged plants was analyzed by RNA-Seq. Seven hundred and ninety four genes were up-regulated and 1018 genes were down-regulated in waterlogged cotton leaves compared with non-waterlogged control. The differentially expressed genes were mainly related to photosynthesis, nitrogen metabolism, starch and sucrose metabolism, glycolysis and plant hormone signal transduction. KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis indicated that most genes related to flavonoid biosynthesis, oxidative phosphorylation, amino acid metabolism and biosynthesis as well as circadian rhythm pathways were differently expressed. Waterlogging increased the expression of anaerobic fermentation related genes, such as alcohol dehydrogenase (ADH), but decreased the leaf chlorophyll concentration and photosynthesis by down-regulating the expression of photosynthesis related genes. Many genes related to plant hormones and transcription factors were differently expressed under waterlogging stress. Most of the ethylene related genes and ethylene-responsive factor-type transcription factors were up-regulated under water-logging stress, suggesting that ethylene may play key roles in the survival of cotton under waterlogging stress.

Gene expression profiles deciphering leaf senescence variation between early- and late-senescence cotton lines.

Leaf senescence varies greatly among genotypes of cotton (Gossypium hirsutium L), possibly due to the different expression of senescence-related genes. To determine genes involved in leaf senescence, we performed genome-wide transcriptional profiling of the main-stem leaves of an early- (K1) and a late-senescence (K2) cotton line at 110 day after planting (DAP) using the Solexa technology. The profiling analysis indicated that 1132 genes were up-regulated and 455 genes down-regulated in K1 compared with K2 at 110 DAP. The Solexa data were highly consistent with, and thus were validated by those from real-time quantitative PCR (RT-PCR). Most of the genes related to photosynthesis, anabolism of carbohydrates and other biomolecules were down-regulated, but those for catabolism of proteins, nucleic acids, lipids and nutrient recycling were mostly up-regulated in K1 compared with K2. Fifty-one differently expressed hormone-related genes were identified, of which 5 ethylene, 3 brassinosteroid (BR), 5 JA, 18 auxin, 8 GA and 1 ABA related genes were up-regulated in K1 compared with K2, indicating that these hormone-related genes might play crucial roles in early senescence of K1 leaves. Many differently expressed transcription factor (TF) genes were identified and 11 NAC and 8 WRKY TF genes were up-regulated in K1 compared with K2, suggesting that TF genes, especially NAC and WRKY genes were involved in early senescence of K1 leaves. Genotypic variation in leaf senescence was attributed to differently expressed genes, particularly hormone-related and TF genes.

[Effects of fertilization on cotton growth and nitrogen use efficiency under salinity stress].

Cotton (Gossypium hirsutum) was raised at different salinity levels (0, 0.15% and 0.30%) by irrigating with fresh- or sea-water. The effects of fertilization (N, NK, NP and NPK) on plant growth, nitrogen (N) uptake and N use efficiency were studied. The results showed that salinity and fertilization both affected the biomass, agronomic N use efficiency, N bioavailability and nitrogen accumulation of plants, and significant interaction was observed between the two factors. Fertilization could improve N use efficiency and nitrogen accumulation of plants under salinity stress, and significantly promoted the cotton yield. Among the fertilization treatments, N combined with P and K had the best effect. The benefit of fertilization was better under low salinity (0.15%) than under moderate salinity (0.3%).