Understanding antioxidant mechanisms for heat stress is important for improving heat tolerance in cool-season plant species. The objective of this study was to identify antioxidant enzymes associated with cultivar variations in heat tolerance in kentucky bluegrass (Poa pratensis) by comparing heat responses of activity and isoforms of antioxidant enzymes in two cultivars contrasting in heat tolerance. Plants of heat-tolerant ‘Eagleton’ and heat-sensitive ‘Brilliant’ were exposed to 20 °C (control) or 40 °C (heat stress) for 28 days in growth chambers. Chlorophyll (Chl) a content remained unchanged and Chl b content increased in ‘Eagleton’, while both of them decreased in ‘Brilliant’, and by 28 days, ‘Eagleton’ had significantly higher Chl a and b content than ‘Brilliant’. The activities of superoxide dismutase (SOD) were significantly higher in ‘Eagleton’ than in ‘Brilliant’ by 28 days of heat stress. An isozyme SOD2 was induced early during heat stress in ‘Eagleton’, while isozyme SOD3 degraded, to a lesser extent in ‘Eagleton’ than in ‘Brilliant’. Catalase (CAT) activity significantly increased in ‘Brilliant’ but remained constant in ‘Eagleton’, and ‘Brilliant’ had a significantly higher CAT activity and isozyme CAT1 than ‘Eagleton’ during heat stress. Significant increases in ascorbate peroxidase (APX) activities occurred under heat stress, to a greater extent in ‘Eagleton’, whereas isozymes did not exhibit difference between cultivars. Guaiacol-peroxidase (POD) activity declined during heat stress in both cultivars. The intensity of POD isozymes in ‘Brilliant’ remained constant, while ‘Eagleton’ showed a transient increases in POD1 at 7 days of heat stress. Our results indicated that antioxidant defense mechanisms for heat tolerance in kentucky bluegrass could be mainly associated with changes in activity and forms of isozymes of SOD for O2 scavenging and APX activity for H2O2 scavenging under heat stress.
Mahalaxmi Veerasamy, Yali He and Bingru Huang
Heat stress induces leaf senescence and causes changes in protein metabolism. The objective of this study was to investigate effects of exogenous application of a synthetic form of cytokinin, zeatin riboside (ZR), on protein metabolism associated with leaf senescence under heat stress for a cool-season grass species. Creeping bentgrass (Agrostis stolonifera L.) (cv. Penncross) plants were exposed to optimum temperature control (20/15 °C, day/night) and heat stress (35/30 °C) in growth chambers. Before heat stress treatments, foliage was sprayed with 10 μmol ZR or water (untreated) for 3 days and then once per week during 35 days of heat stress. Leaf chlorophyll content, photochemical efficiency (Fv/Fm), and soluble protein content declined, whereas protease activity increased during heat stress. Treatments with ZR helped maintain higher leaf chlorophyll content, Fv/Fm, and soluble protein content under heat stress. Protease activity in ZR-treated plants was lower than that of untreated plants. Zeatin riboside-treated plants had less severe degradation of ribulose-1,5-bisphosphate carboxylase proteins than untreated plants exposed to heat stress. In addition, ZR treatment upregulated the expression of 32- and 57-kDa proteins under heat stress conditions. These results demonstrated that the exogenous application of ZR ameliorated the negative effects of heat stress, as manifested by suppression or delay of leaf senescence. Cytokinins may have helped to alleviate heat stress injury, probably by slowing down the action of protease and by induction or upregulation of heat-shock proteins.
Yali He, Xiaozhong Liu and Bingru Huang
Various physiological processes may deteriorate in response to increasing temperatures, contributing to the decline in turf quality for cool-season turfgrasses during heat stress. This study was performed to investigate metabolic changes (membrane lipid peroxidation, total protein content, amino acid content, and protease activity) associated with turf quality decline for creeping bentgrass (Agrostis stolonifera Huds.) in response to gradually increasing temperatures for a short duration and prolonged exposure to lethally high temperature. Plants were subjected to increasing temperatures of 20, 25, 30, 35, and 40 °C for 7 days at each level of temperature [gradual heat stress (GHS)] or exposed to high temperature of 40 °C for 28 days [prolonged heat stress (PHS)] in growth chambers. During the GHS treatment, significant decline in turf quality occurred when plants were exposed to 30 °C for 7 days; simultaneously, malondialdehyde (MDA) content increased and total protein content in shoots decreased significantly compared to those at 20 °C. Protease activity increased at 25 °C and then decreased as temperature was elevated from 30 to 40 °C during the GHS treatment. Amino acid content decreased under GHS, beginning at 25 °C. Under the PHS treatment, turf quality declined and MDA content increased significantly, beginning at 14 days of PHS, while total protein content decreased at 7 days of PHS. Protease activity and amino acid content increased at 7 days of PHS, and then declined with longer stress duration. Our results indicated that protease activity, and amino acid and total protein content were more responsive to GHS or PHS than that of lipid peroxidation and turf quality. Changes in metabolic parameters of protease activity, amino acid and total protein content, and lipid peroxidation may contribute to leaf senescence and poor turf performance under severe or prolonged heat stress conditions for creeping bentgrass.
Yali He, Xiaozhong Liu and Bingru Huang
The acclimation of plants to moderately high temperature plays an important role in inducing plant tolerance to subsequent lethal high temperatures. This study was performed to investigate the effects of heat acclimation and sudden heat stress on protein synthesis and degradation in creeping bentgrass (Agrostis palustris Huds.). Plants of the cultivar Penncross were subjected to two temperature regimes in growth chambers: 1) heat acclimation—plants were exposed to a gradual increase in temperatures from 20 to 25, 30, and 35 °C for 7 days at each temperature level before being exposed to 40 °C for 28 days; and 2) sudden heat stress (nonacclimation)—plants were directly exposed to 40 °C for 28 days from 20 °C without acclimation through the gradual increase in temperatures. Heat acclimation increased plant tolerance to subsequent heat stress, as demonstrated by lower electrolyte leakage (relative EL) in leaves of heat-acclimated plants compared to nonacclimated plants at 40 °C. Heat acclimation induced expression of some heat shock proteins (HSPs), 57 and 54 kDa, detected in a salt-soluble form (cystoplasmic proteins), which were not present in unacclimated plants under heat stress. However, HSPs of 23, 36, and 66 kDa were induced by both sudden and gradual exposure to heat stress. In general, total protein content decreased under both heat acclimation and sudden heat stress. Cystoplasmic proteins was more sensitive to increasing temperatures, with a significant decline initiated at 25 °C, while sodium dodecyl sulphate (SDS)-soluble (membrane) protein content did not decrease significantly until temperature was elevated to 30 °C. The results demonstrated that both a gradual increase in temperature and sudden heat stress caused protein degradation and also induced expression of newly synthesized HSPs. Our results suggested that the induction of new HSPs during heat acclimation might be associated with the enhanced thermotolerance of creeping bentgrass, although direct correlation of these two factors is yet to be determined. This study also indicated that protein degradation could be associated with heat injury during either gradual increases in temperature or sudden heat stress.
Mengmeng Zhang, He Huang, Qing Wang and Silan Dai
Conventional crossbreeding remains an effective technique for chrysanthemum (Chrysanthemum ×morifolium Ramat.) breeding. However, there are always many problems when breeding chrysanthemum because of its complex genetic background, such as difficulty matching parents, selecting superior hybrid progenies, quantitatively describing certain target traits, and evaluating breeding results. A recent mathematical analysis method is an effective method for evaluating plant breeding progress. In this study, we used 505 multiflora chrysanthemum germplasm resources as test materials; we divided the flowering time into five groups using a grading analysis method, including extremely early group (genotypes that flowered when daylength was longer than 13.5 hours), early group (genotypes that flowered when daylength was 13.5–12.0 hours), medium group (genotypes that flowered when daylength was 12.0–11.0 hours), late group (genotypes that flowered when daylength was 11.0–10.0 hours), and extremely late group (genotypes that flowered when daylength was shorter than 10.0 hours). Moreover, the breeding objective was to breed early-flowering genotypes. Using 15 phenotypic characters as evaluation factors, 37 excellent genotypes, including four early-flowering genotypes, were screened out from the aforementioned resources according to an analytic hierarchy process (AHP) and weighting of the gray relational grade. We selected one early-flowering genotype and eight medium-flowering genotypes from these 37 genotypes and matched six hybridized combinations based on the genetic distance between genotypes calculated by the Q cluster analysis method. We used a comprehensive evaluation method combining AHP and the gray relational analysis (GRA) method for the evaluation of 367 progenies. Moreover, we screened out 52 superior hybrids, including 36 early-flowering hybrids. The results of this study demonstrate that the mathematical analysis method is an immensely effective method to breed new cultivars of early-flowering multiflora chrysanthemum. This study also provides an effective method to define and improve the flowering time of other cultivated plants.
Yu Liu, Miao He, Fengli Dong, Yingjie Cai, Wenjie Gao, Yunwei Zhou, He Huang and Silan Dai
The NAC transcription factor is a peculiar kind of transcription factor in plants. Transcription factors are involved in the expression of plant genes under different conditions, and they play a crucial role in plant response to various biotic and abiotic stress. We transferred the ClNAC9 gene into Chrysanthemum grandiflora ‘niu9717’ by Agrobacterium tumefaciens–mediated transformation. The results of kanamycin-resistant screening, polymerase chain reaction (PCR) detection, and Northern blot analysis proved that the target gene had been integrated into the genome of the target plants. Wild-type (WT) plants and transgenic plants were treated with different concentrations of NaCl, NaHCO3, and drought stress, and physiological indexes, such as antioxidant system activity (superoxide dismutase, peroxidase, catalase), malondialdehyde accumulation, and leaf relative water content, were measured. We also observed changes in plant morphology. The physiological indexes’ changing range and extreme values suggested that transgenic plants’ resistance to salinity, alkali, and drought stress was significantly higher than WT plants. Transgenic plant growth was less inhibited compared with WT plants, indicating that the ClNAC9 gene increased the resistance of transgenic plants under the stress of salinization, alkalization, and drought.
Peitao Lü, Xinmin Huang, Hongmei Li, Jiping Liu, Shenggen He, Daryl C. Joyce and Zhaoqi Zhang
In studying the postharvest water relations of cut flowers, researchers aim to determine rates of water uptake and water loss along with changes in fresh weight. An automatic apparatus was devised for continuous monitoring of these indices. The novel apparatus consists of two balances automatically recording mass at a relatively high data acquisition rate (min−1), a personal computer, two containers, and plastic tubing. The apparatus is accurate, labor-saving, and real-time. It enabled dynamic synchronous recording of water uptake as well as fresh weight of the cut flower stem, from which precise water uptake loss rates during vase life can be accurately determined. Rates of water uptake and water loss of individual cut rose (Rosa hybrida cv. Movie Star) stems were measured using the apparatus under alternating 12-h light and dark periods. Both water uptake and water loss rates fluctuated with the light to dark shift over 120 h of observation. Stem fresh weight increased rapidly over the first 40 h of vase period and decreased gradually thereafter. Cut lily (Lilium hybrida cv. Yellow Overlord) stems showed similar trends in water uptake and water loss rate to cut rose stems. The accuracy and sensitivity of the new apparatus was validated by comparison with manual weighing using a balance at 2-h intervals under alternating 12-h light and dark periods over 108 h. The apparatus described here constitutes a suitable method for direct measurement of water uptake and fresh weight, including capturing relatively rapid water balance responses to changes in the postharvest environment.
Xiaotao Ding, Yuping Jiang, Dafeng Hui, Lizhong He, Danfeng Huang, Jizhu Yu and Qiang Zhou
Adequate greenhouse environmental management is very important for improving resource use efficiency and increasing vegetable yield. The objective of this study was to explore suitable climate and cultivation management for cucumber to achieve high yield and build optimal yield models in semi-closed greenhouses. A fruit cucumber cultivar Deltastar was grown over 4 years in greenhouse and weekly data of yields (mean, highest and lowest) and environmental variables, including total radiation, air temperature, relative humidity, and carbon dioxide (CO2) concentration were collected. Regression analyses were applied to develop the relationships and build best regression models of yields with environmental variables using the first 2 years of data. Data collected in years 3 and 4 were used for model validation. Results showed that total radiation, nutrient, temperature, CO2 concentration, and average nighttime relative humidity had significant correlations with cucumber yields. The best regression models fit the mean, lowest, and highest yields very well with R 2 values of 0.67, 0.66, and 0.64, respectively. Total radiation and air temperature had the most significant contributions to the variations of the yields. Our results of this study provide useful information for improving greenhouse climate management and yield forecast in semi-closed greenhouses.
He Huang, Yuting Liu, Ya Pu, Mi Zhang and Silan Dai
Plant growth and development are significantly affected by salt stress. Chrysanthemum lavandulifolium is a halophyte species and one of the ancestors of chrysanthemum (C. ×morifolium). Understanding how this species tolerates salt stress could provide vital insight for clarifying the salt response systems of higher plants, and chrysanthemum-breeding programs could be improved. In this study, salt tolerance was compared among C. lavandulifolium and three chrysanthemum cultivars by physiological experiments, among which C. lavandulifolium and Jinba displayed better tolerance to salt stress than the other two cultivars, whereas Xueshan was a salt-sensitive cultivar. Using the transcriptome database of C. lavandulifolium as a reference, we used digital gene expression technology to analyze the global gene expression changes in C. lavandulifolium seedlings treated with 200 mm NaCl for 12 hours compared with seedlings cultured in normal conditions. In total, 2254 differentially expressed genes (DEGs), including 1418 up-regulated and 836 down-regulated genes, were identified. These DEGs were significantly enriched in 35 gene ontology terms and 29 Kyoto Encyclopedia of Genes and Genomes pathways. Genes related to signal transduction, ion transport, proline biosynthesis, reactive oxygen species scavenging systems, and flavonoid biosynthesis pathways were relevant to the salt tolerance of C. lavandulifolium. Furthermore, comparative gene expression analysis was conducted using reverse transcription polymerase chain reaction to compare the transcriptional levels of significantly up-regulated DEGs in C. lavandulifolium and the salt-sensitive cultivar Xueshan, and species-specific differences were observed. The analysis of one of the DEGs, ClAKT, an important K+ transport gene, was found to enable transgenic Arabidopsis thaliana to absorb K+ and efflux Na+ under salt stress and to absorb K+ under drought stress. The present study investigated potential genes and pathways involved in salt tolerance in C. lavandulifolium and provided a hereditary resource for the confinement of genes and pathways responsible for salt tolerance in this species. This study provided a valuable source of reference genes for chrysanthemum cultivar transgenesis breeding.
Zhi Quan, Bin Huang, Caiyan Lu, Yi Shi, Yanhong Cao, Yongzhuang Wang, Chuanrui He, Guangyu Chi, Jian Ma and Xin Chen
Much nitrogen (N) is lost in high-input protected cropping systems mainly via leaching of not only nitrate-N but also extractable organic N (EON), but the role of EON in this process is poorly appreciated. A consecutive 3-year plot experiment was conducted to investigate the impact of co-application of manures with chemical N fertilizer on N accumulation and loss in a greenhouse soil rotationally planted with cucumber or tomato and lettuce. Application of manures significantly enhanced the average contents and stocks of NO3 −-N, EON, and total N (TN) in 0- to 60-cm soil layer, although EON accumulated within growing season, while NO3 −-N accumulated with fluctuation, and TN accumulated gradually throughout the 3-year experiment. With application rate at 120 or 180 t dry manures per hectare per 3 years, the corresponding apparent N surplus was 2710 or 3924 kg⋅ha−1 per 3 years. Due to little increase of biomass N uptakes during vegetable seasons, the accumulated N in soil profile would be a potential loss source, largely via leaching of both nitrate and EON. Application of manures slowed soil acidification but intensified secondary salinization of the greenhouse soil. Considering the manures-induced high soil N accumulation and loss, well-balanced evaluation of the role of manures in high-input agricultural ecosystems is needed.