Xiaoli Tian

Overexpression of the AtLOS5 gene increased abscisic acid level and drought tolerance in transgenic cotton

Drought is the major environmental stress that limits cotton (Gossypium hirsutum L.) production worldwide. LOS5/ABA3 (LOS5) encodes a molybdenum co-factor and is essential for activating aldehyde oxidase, which is involved in abscisic acid (ABA) biosynthesis. In this study, a LOS5 cDNA of Arabidopsis thaliana was overexpressed in cotton cultivar Zhongmiansuo35 (Z35) by Agrobacterium tumefaciens-mediated transformation. The transformation and overexpression of AtLOS5 were assessed by PCR and RT-PCR analysis. Detached shoots of transgenic cotton showed slower transpirational water loss than those of Z35. When pot-grown 6-week-old seedlings were withheld from watering for 3 d, transgenic cotton accumulated 25% more endogenous ABA and about 20% more proline than Z35 plants. The transgenic plants also showed increased expression of some drought-responding genes such as P5CS and RD22, and enhanced activity of antioxidant enzymes such as superoxide dismutase, peroxidase, and ascorbate peroxidase. Their membrane integrity was considerably improved under water stress, as indicated by reduced malondialdehyde content and electrolyte leakage relative to control plants. When the pot-grown plants were subjected to deficit irrigation for 8 weeks (watering to 50% of field capacity), transgenic plants showed a 13% increase in fresh weight than the wild type under the same drought condition. These results suggest that the AtLOS5 transgenic cotton plants acquired a better drought tolerance through enhanced ABA production and ABA-induced physiological regulations.

Differential responses of conventional and Bt-transgenic cotton to potassium deficiency

ABSTRACT Bacillus thuringensis (Bt) transgenic (insect-resistant) cotton cultivars senesce prematurely under potassium (K+) deficiency, more often than conventional cultivars, in the North China Plain. To verify if Bt-transgenic cotton was more susceptible to K+ deficit, two transgenic cultivars, ‘CCRI 41’ and ‘DP 99B’, and two conventional cultivars, ‘CCRI 35’ and ‘CCRI 36’, selected from widely used cultivars in China, were used in a seedling hydroponic study. The culture solution K+ concentration was 0.5 mM for high K+ and 0.02 mM for low K+ conditions. Seedlings of all four cultivars accumulated more dry matter and K+ when grown at high K+ than low K+ conditions. However, under low K+ condition, the dry weight and K+ content of Bt-transgenic cultivars CCRI 41 and DP 99B were lower than those of the conventional cultivars CCRI 36 and CCRI 35. The results indicated that Bt-transgenic cultivars CCRI 41 and DP 99B were more sensitive to K+ deficiency than conventional cultivars CCRI 36 and CCRI 35, which could be the reason for premature senescence symptoms observed from fields of Bt-transgenic cotton under K+ deficiency. Seedlings of all four cultivars had a higher K+ use efficiency (KUE) under low K+ than high K+ conditions, but the KUE did not account for the differential responses between Bt-transgenic and conventional cultivars at the low K+ concentration. The K+ depletion results did not reveal the mechanism for the above differential responses in Vmax and Km of the seedlings either. Further experiments with more cultivars are needed to clarify the differential mechanisms in these genotypes.

Genotypic variations in photosynthetic and physiological adjustment to potassium deficiency in cotton (Gossypium hirsutum)

A hydroponic culture experiment was conducted to determine genotypic variation in photosynthetic rate and the associated physiological changes in response to potassium (K) deficiency in cotton (Gossypium hirsutum L.) seedlings with contrasting two cotton cultivars in K efficiency. The K-efficient Liaomian18 produced 66.7% more biomass than the K-inefficient NuCOTN99(B) under K deficiency, despite their similar biomass under K sufficiency. Compared with NuCOTN99(B), Liaomian18 showed 19.4% higher net photosynthetic rate (P(n), per unit leaf area) under K deficient solutions and this was associated with higher photochemical efficiency and faster export of soluble sugars from the phloem. The lower net P(n) of NuCOTN99(B) was attributed to higher capacity for nitrate assimilation and lower export of soluble sugars. Furthermore, NuCOTN99(B) showed 38.4% greater ETR/P(n) than Liaomian18 under K deficiency, indicating that more electrons were driven to other sinks. Higher superoxide dismutase (SOD) and lower catalase (CAT) and ascorbate peroxidase (APX) activities resulted in higher levels of reactive oxygen species (ROS; e.g. O(2)(-)and H(2)O(2)) in NuCOTN99(B) relative to Liaomian18. Thus, the K inefficiency of NuCOTN99(B), indicated by lower biomass and net P(n) under K deficiency, was associated with excessively high nitrogen assimilation, lower export of carbon assimilates, and greater ROS accumulation in the leaf.

Mechanism of phytohormone involvement in feedback regulation of cotton leaf senescence induced by potassium deficiency

To elucidate the phytohormonal basis of the feedback regulation of leaf senescence induced by potassium (K) deficiency in cotton (Gossypium hirsutum L.), two cultivars contrasting in sensitivity to K deficiency were self- and reciprocally grafted hypocotyl-to-hypocotyl, using standard grafting (one scion grafted onto one rootstock), Y grafting (two scions grafted onto one rootstock), and inverted Y grafting (one scion grafted onto two rootstocks) at the seedling stage. K deficiency (0.03mM for standard and Y grafting, and 0.01mM for inverted Y grafting) increased the root abscisic acid (ABA) concentration by 1.6- to 3.1-fold and xylem ABA delivery rates by 1.8- to 4.6-fold. The K deficiency also decreased the delivery rates of xylem cytokinins [CKs; including the zeatin riboside (ZR) and isopentenyl adenosine (iPA) type] by 29–65% and leaf CK concentration by 16–57%. The leaf ABA concentration and xylem ABA deliveries were consistently greater in CCRI41 (more sensitive to K deficiency) than in SCRC22 (less sensitive to K deficiency) scions under K deficiency, and ZR- and iPA-type levels were consistently lower in the former than in the latter, irrespective of rootstock cultivar or grafting type, indicating that cotton shoot influences the levels of ABA and CKs in leaves and xylem sap. Because the scions had little influence on phytohormone levels in the roots (rootstocks) of all three types of grafts and rootstock xylem sap (collected below the graft union) of Y and inverted Y grafts, it appears that the site for basipetal feedback signal(s) involved in the regulation of xylem phytohormones is the hypocotyl of cotton seedlings. Also, the target of this feedback signal(s) is more likely to be the changes in xylem phytohormones within tissues of the hypocotyl rather than the export of phytohormones from the roots.

Effects of Coronatine on Growth, Gas Exchange Traits, Chlorophyll Content, Antioxidant Enzymes and Lipid Peroxidation in Maize (Zea mays L.) Seedlings under Simulated Drought Stress

Abstract Coronatine is a phytotoxin that affects the accumulation of defence-related metabolites in plants but information on how its effects may be mediated by environmental stress is scanty. An experiment was carried out to determine the changes in growth, gas exchange, relative water content, chlorophyll (Chl) content, antioxidant enzymes and lipid peroxidation in maize (Zea mays L., var. ‘Nongda 3138’) seedlings treated with coronatine under simulated drought stress. Seedlings raised hydroponically in a growth chamber with simulated drought for 8d (long-period drought) or 3d (short-period drought) were treated with or without coronatine at the three-leaf stage. Under the drought condition, treated with coronatine significantly increased the fresh weight and relative water content in leaves of seedling leaves. The increase was accompanied by increased rates of photosynthesis and transpiration, and the maintenance of Chl pigments. Coronatine had no effects on catalase (CAT), guaiacol peroxidase (POD) and glutathione reductase (GR) under normal condition, but it significantly enhanced activities of CAT, POD and GR in stressed seedlings under the long-period drought treatment. Under the short-period drought treatment, the POD and GR activity in the seedlings treated with coronatine were much higher than in those not treated. Malondialdehyde (MDA) increased sharply under drought condition, but treatment with coronatine significantly reduced it by 15%. The total Chl content of leaves under the drought condition was markedly increased by the treatment with coronatine. Seedlings subjected to a short-period drought had reduced water content, but recovered fairly well by the treatment with coronatine with negligible effects on most physiological and biochemical processes. The application of coronatine alleviated the drought stress in maize seedlings and enhanced their tolerance of water stress through changes in physiological and anti-oxidant enzyme activities.

GENOTYPIC VARIATIONS IN POTASSIUM UPTAKE AND UTILIZATION IN COTTON

Genotypic differences in potassium (K) uptake and utilization were compared for eight cotton cultivars in growth chamber and field experiments. Four of the cultivars (‘SGK3’, ‘SCRC18’, ‘SCRC21’ and ‘SCRC22’) typically produce lower dry mass and the other four (‘Nannong8’, ‘Xiangza2’, ‘Xinluzao12’ and ‘Xiangza3’) produce greater dry mass in K-deficient solution (0.02 mM). The mean dry weight of seedlings (five-leaf stage) of cultivars with greater biomass was 155% higher than that of cultivars with lower biomass yield under K deficiency. However, all the genotypes had similar dry matter yields in K-sufficient solution (2.5 mM). Thus, the four cultivars with superior biomass yield under low K medium may be described as K efficient cultivars while the inferior cultivars may be described as K inefficient. Although seeds of the studied cultivars originated from different research institutes or seed companies, there were little differences in seed K content among them, irrespective of their K efficiency. Consequently, there were no significant differences in K accumulation in seedlings (4 d after germination in a K-free sand medium) just before transferring to nutrient solutions. However, the K efficient genotypes, on average, accumulated twice as much K at 21 d after transferring to K-deficient solution (0.02 mM). A much larger root system as well as a slightly higher uptake rate (K uptake per unit of root dry weight) may have contributed to the higher net K uptake by the K efficient cultivars. In addition, the K efficiency ratio (dry mass produced per unit of K accumulated) and K utilization efficiency (dry mass produced per unit of K concentration) of the K efficient cultivars exceeded those of the K inefficient genotypes by 29% and 234%, respectively, under K deficiency. On average, the K efficient cultivars produced 59% more potential economic yield (dry weight of all reproductive organs) under field conditions even with available soil K at obviously deficient level (60 mg kg−1). We noted especially that the four K inefficient cultivars studied were all transgenic insect-resistant cotton, suggesting that the introduction of foreign genes (Bt and CpTI) may affect the K use efficiency of cotton.

Cotton shoot plays a major role in mediating senescence induced by potassium deficiency.

The objective of this study was to determine the roles of shoot and root in the regulation of premature leaf senescence induced by potassium (K) deficiency in cotton (Gossypium hirsutum L.). Two contrasting cultivars (CCRI41, more sensitive to K deficiency; and SCRC22, a less sensitive cultivar) were selected for self- and reciprocal-grafting, using standard grafting (one scion/one rootstock), Y grafting (two scions/one rootstock) and inverted Y grafting (one scion/two rootstocks) at the seedling stage. Standard grafting was studied in the field in 2007 and 2008. There were no obvious differences in senescence between CCRI41 and SCRC22 scions while supplied with sufficient K. However, SCRC22 scions showed significantly greater K content, SPAD values (chlorophyll content), soluble protein content and net photosynthetic rates than CCRI41 scions while grown in K deficient solution or soil, regardless of rootstock cultivars, grafting types, growth stage and growth conditions. Also, SCRC22 scions had greater yield and less variation in boll weight either between upper- and lower sympodials, or between proximal and distal fruit positions from the main stem in the field under K deficiency, probably owing to reduced leaf senescence. Although the effect of rootstocks on leaf senescence under K deficiency was significant in some cases, the scion cultivars explained the highest percentage of variations within grafting treatments. The shoot-to-root feedback signal(s), rather than high shoot demand for K nutrition, was involved in the shoot regulation of premature senescence in cotton plants, achieved possibly by altering root K uptake.

The effect of mepiquat chloride on elongation of cotton (Gossypium hirsutum L.) internode is associated with low concentration of gibberellic acid.

The growth regulator mepiquat chloride (MC) is globally used in cotton (Gossypium hirsutum L.) canopy manipulation to avoid excess growth and yield loss. However, little information is available as to whether the modification of plant architecture by MC is related to alterations in gibberellic acid (GA) metabolism and signaling. Here, the role of GA metabolism and signaling was investigated in cotton seedlings treated with MC. The MC significantly decreased endogenous GA3 and GA4 levels in the elongating internode, which inhibited cell elongation by downregulating GhEXP and GhXTH2, and then reducing plant height. Biosynthetic and metabolic genes of GA were markedly suppressed within 2-10d of MC treatment, which also downregulated the expression of DELLA-like genes. A remarkable feedback regulation was observed at the early stage of MC treatment when GA biosynthetic and metabolic genes expression was evidently upregulated. Mepiquat chloride action was controlled by temporal translocation and spatial accumulation which regulated GA biosynthesis and signal expression for maintaining GA homeostasis. The results suggested that MC application could reduce endogenous GA levels in cotton through controlled GA biosynthetic and metabolic genes expression, which might inhibit cell elongation, thereby shortening the internode and reducing plant height.

Coronatine Alleviates Water Deficiency Stress on Winter Wheat Seedlings

With the aim to determine whether coronatine (COR) alleviates drought stress on wheat, two winter wheat (Triticum aestivum L.) cultivars, ChangWu134 (drought-tolerant) and Shan253 (drought-sensitive) were studied under hydroponic conditions. Seedlings at the three-leaf stage were cultured in a Hoagland solution containing COR at 0.1 microM for 24 h, and then exposed to 20% polyethylene glycol 6000 (PEG-6000). Under simulated drought (SD), COR increased the dry weight of shoots and roots of the two cultivars significantly; the root/shoot ratio also increased by 30% for Shan253 and 40% for ChangWu134. Both cultivars treated with COR under SD (0.1COR+PEG) maintained significantly higher relative water content, photosynthesis, transpiration, intercellular concentration of CO(2) and stomatal conductance in leaves than those not treated with PEG. Under drought, COR significantly decreased the relative conductivity and malondialdehyde production, and the loss of 1,1-diphenyl-2-picrylhydrazyl scavenging activity in leaves was significantly alleviated in COR-treated plants. The activity of peroxidase, catalase, glutathione reductase and ascorbate peroxidase were adversely affected by drought. Leaves of plants treated with COR under drought produced less abscisic acid (ABA) than those not treated. Thus, COR might alleviate drought effects on wheat by reducing active oxygen species production, activating antioxidant enzymes and changing the ABA level.

Effects of Potassium Deficiency on Root Growth of Cotton Seedlings and Its Physiological Mechanisms

Abstract For the purpose of dissecting the mechanism of root growth in response to potassium (K) deficiency in cotton (Gossypium hirsutum L.), young seedlings of NuCOTN99B grown in half-strength modified Hoaglands solution with low K nutrient (0.05 mmol L−1) were investigated for the root configuration, content of endogenous free indole acetic acid (IAA), and amount of ethylene released from the roots 4 d after treatment. Compared with the treatment with moderate K nutrient (0.50 mmol L−1, control), the K deficient treatment significantly inhibited root length and the formation of lateral roots. The reduced lateral roots mainly resulted from the shortened branched root zone, and there was no change in the lateral root density. Under K deficient condition, the greatest reductions for root length, total root surface area, and root volume occurred in fine roots (0.05 mm ≤ diameter