Calcium Signaling is Involved in Negative Phototropism of Rice Seminal Roots

Calcium ions （Ca2＋） act as an intracellular second messenger and affect nearly all aspects of cellular life. They are functioned by interacting with polar auxin transport, and the negative phototropism of plant roots is caused by the transport of auxin from the irradiated side to the shaded side of the roots. To clarify the role of calcium signaling in the modulation of rice root negative phototropism, as well as the relationship between polar auxin transport and calcium signaling, calcium signaling reagents were used to treat rice seminal roots which were cultivated in hydroculture and unilaterally illuminated at an intensity of 100-200 pmol/（m2.s） for 24 h. Negative phototropism curvature and growth rate of rice roots were both promoted by exogenous CaCI2 lower than 100 pmol/L, but inhibited by calcium channel blockers （verapamil and LaCI3）, calcineurin inhibitor （chlorpromazine, CPZ）, and polar auxin transport inhibitor （N-l-naphthylphthalamic acid, NPA）. Roots stopped growing and negative phototropism disappeared when the concentrations increased to 100 pmol/L verapamil, 12.500 ~Jmol/L LaCI3, 60 pmol/L CPZ, and 6 pmol/L NPA. Moreover, 100 pmol/L CaCI2 could relieve the inhibition of LaCI3, verapamil and NPA. The enhanced negative phototropism curvature was caused by the transportation of more auxin from the irradiated side to the shaded side in the presence of exogenous Ca2＋. Calcium signaling plays a key role as a second messenger in the process of light signal regulation of rice root growth and negative phototropism.     

Effect of Indoleacetic acid （IAA） on the Negative Phototropism of Rice Root

To explore the effects of IAA on negative phototropism of rice (Oryza sativa L) root, agar block containing IAA was unilaterally applied on root tip to examine the phototropic response of root to exogenous IAA, and microstructure of the bending part was observed with an optical microscope. The growth of seminal roots could be regulated by exogenous IAA as well as light,as a result the root bent towards the site treated, causing asymmetric growth of the root cells at the elongation zone and consequently bending growth. IAA concentration in the shaded side of adventitious root increased much greater at 1.5 h after the start of irradiation. The unequal lateral IAA distribution can be concluded to be the main cause for negative phototropism of rice root.     

Discussion on Photoreceptor for Negative Phototropism in Rice Roots

To properly explore the photoreceptor for the negative phototropism in rice (Oryza sativa L.) root, lights with different wavelengths were applied to investigate the effect of light quality on phototropic bending. The phototropic bending could be induced prominently by blue/ultraviolet light, whereas not by red or far-red light. The absorption spectrum of the extracted solution from rice root cap had two peaks at 350 nm and 450 nm, respectively, and the molecular weight of the 120 kD protein in the root cap under unilateral light was larger than that under the dark. It suggested that the blue light receptor might be the photoreceptor for the negative phototropism in rice root.     

Physiological and Proteomic Analyses Reveal Effects of Putrescine-Alleviated Aluminum Toxicity in Rice Roots

The effects of putrescine on improving rice growth under aluminum(Al) toxicity conditions have been previously demonstrated, however, the underlying mechanism remains unclear. In this study, treatment with 50 μmol/L Al significantly decreased rice root growth and whole rice dry weight, inhibited Ca~(2+) uptake, decreased ATP synthesis, and increased Al, H_2O_2 and malondialdehyde(MDA) contents, whereas the application of putrescine mitigated these negative effects. Putrescine increased root growth and total dry weight of rice, reduced total Al content, decreased H_2O_2 and MDA contents by increasing antioxidant enzyme(superoxide dismutase, peroxidase, catalase and glutathione S-transferase) activities, increased Ca~(2+) uptake and energy production. Proteomic analyses using data-independent acquisition successfully identified 7 934 proteins, and 59 representative proteins exhibiting fold-change values higher than 1.5 were randomly selected. From the results of the proteomic and biochemical analyses, we found that putrescine significantly inhibited ethylene biosynthesis and phosphorus uptake in rice roots, increased pectin methylation, decreased pectin content and apoplastic Al deposition in rice roots. Putrescine also alleviated Al toxicity by repairing damaged DNA and increasing the proteins involved in maintaining plasma membrane integrity and normal cell proliferation. These findings improve our understanding of how putrescine affects the rice response to Al toxicity, which will facilitate further studies on environmental protection, crop safety, innovations in rice performance and real-world production.     

Purification and Identification of Glutathione S-transferase in Rice Root under Cadmium Stress

Cadmium (Cd) contamination in paddy soils poses a serious threat to the production and quality of rice.Among various biochemical processes related to Cd detoxification in rice,glutathione S-transferase (GST) plays an important role,catalyzing Cd complexation with glutathione (GSH) and scavenging reactive oxygen species (ROS) in cells.In this study,a hydroponic experiment was conducted to investigate the response of GST isozymes in rice roots upon Cd exposure.Results showed that the GST activity in rice roots was clearly enhanced by 50 μmol/L Cd treatment for 7 d.The GST isozymes were purified by ammonium sulphate precipitation,gel filtration chromatography and affinity chromatography.After being separated by SDS-PAGE and visualized by silver staining,GSTU6 was identified by in-gel digestion,MALDI-TOF-MS analysis and peptide mass fingerprint.The results confirm the vital function of tau class rice GST in Cd detoxification.     

Effects of 1,2,4-Trichlorobenzene and Mercury Ion Stress on Ca^2＋ Fluxion and Protein Phosphorylation in Rice

The effects of 5 mg/L 1,2,4-trichlorobenzene (TCB) and 0.1 mmol/L mercury ion (Hg2 ) stresses on Ca2 fluxion and protein phosphorylation in rice seedlings were investigated by isotope exchange kinetics and in vitro phosphorylation assay. The Ca2 absorption in rice leaves and Ca2 transportation from roots to leaves were promoted significantly in response to Hg2 and TCB treatments for 4-48 h. The Ca2 absorption peaks presented in the leaves when the rice seedlings were exposed to Hg2 for 8-12 h or to TCB for 12-24 h. Several Ca2 absorption peaks presented in the roots during rice seedlings being exposed to Hg2 and TCB, and the first Ca2 absorption peak was at 8 h after being exposed to Hg2 and TCB. The result of isotope exchange kinetic analysis confirmed that short-term (8 h) Hg2 and TCB stresses caused Ca2 channels or pumps located on plasmalemma to open transiently. The phosphorylation assay showed that short-term TCB stress enhanced protein phosphorylation in rice roots (TCB treatment for 4-8 h) and leaves (TCB treatment for 4-24 h), and short-term (4-8 h) Hg2 stress also enhanced protein phosphorylation in rice leaves. The enhancement of protein phosphorylation in both roots and leaves corresponded with the first Ca2 absorption peak, which confirmed that the enhancement of protein phosphorylation caused by TCB or Hg2 stress might be partly triggered by the increases of cytosolic calcium. TCB treatment over 12 h inhibited protein phosphorylation in rice roots, which might be partly due to that TCB stress suppressed the protein kinase activity. Whereas, Hg2 treatment inhibited protein phosphorylation in rice roots, and Hg2 treatment over 12 h inhibited protein phosphorylation in rice leaves. This might be attributed to that not only the protein kinase activity, but also the expressions of phosphorylation proteins were restrained by Hg2 stress.     

Responses of Four Rice Varieties to Elevated CO2 and Different Salinity Levels

This study was carried out in 2014 at Isfahan University of Technology, Iran, to evaluate the responses of four rice varieties(Neda, Deylamani, Shiroudi and Domsorkh) to ambient(360 ± 50 μmol/mol) and elevated(700 ± 50 μmol/mol) air carbon dioxide(CO_2) concentrations under four salinity levels(0, 30, 60 and 90 mmol/L Na Cl). There was significant variation among rice varieties in response to elevated CO_2 concentration under the four salinity levels. Under non-saline condition, elevated CO_2 increased the dry weight of Neda, Deylamani and Domsorkh by 8%, 50% and 8%, respectively, but reversely decreased that of Shiroudi by 34%. Increasing CO_2 concentration significantly reduced the negative effects of salinity on Shiroudi, but these effects were even increased in Deylamani and Domsorkh under all the salinity levels and in Neda only under 30 and 60 mmol/L Na Cl. Significant correlations were established between plant dry weight, SPAD value and leaf area under both CO_2 levels. However, this trend was observed only at ambient CO_2 concentration in the presence of soluble carbohydrates. The results revealed the genotype and salinity dependence of the effects of CO_2 concentrations on the rice traits investigated.     

Genetic Analysis and Molecular Mapping of Light-Sensitive Red-Root Mutant in Rice

The light-sensitive red-root mutant, designated as HG1, was newly observed from an indica rice variety, Nankinkodo, when seedlings were grown with roots exposed to natural light. The root color of the mutant began to turn slight-red when the roots were exposed to the light at the intensity of 29 μmol/(m2·s), then turned dark-red at the light intensity of 180 μmol/(m2·s), suggesting that the root color of the mutant was evidently sensitive to light. Furthermore, genetic analysis showed that the character of ...     

钙信号试剂对水稻种子根负向光性生长的影响

 为了探讨水稻根负向光性形成与钙信号传导的关系,以水稻种子根为材料,用不同浓度的钙信号试剂\[氯化钙、钙通道有机阻断剂异搏定、钙通道无机阻断剂氯化镧、钙调素抑制剂氯丙嗪（CPZ）以及生长素极性运输抑制剂NPA\]处理水稻种子根,以1/10 Hoagland培养液为对照,并用100~200 μmol/(m2·s)单侧光照射24 h。实验结果表明,适量的CaCl2使水稻种子根负向光性增强并使其生长加快,负向光性增强是由于外源Ca2+进一步促进生长素从向光侧向背光侧运输引起的,与生长速率加快无关;同时,随着钙通道有机阻断剂、无机阻断剂、钙调素抑制剂以及生长素极性运输抑制剂浓度的上升,水稻根的负向光性及生长均受显著抑制,当异搏定的浓度大于100 μmol/L,LaCl3的浓度大于12.5 μmol/L,负向光性消失,种子根的生长也受到严重的影响;而60 μmol/L钙调素抑制剂氯丙嗪、6 μmol/L 生长素极性运输抑制剂NPA也使得根负向光性消失,根的生长几乎停止;向LaCl3溶液、异搏定溶液以及NPA溶液中添加100 μmol/L CaCl2可使负向光性及生长速率得到不同程度的恢复,表明钙离子作为第二信使系统,与生长素相互作用,参与光信号调控水稻根生长和负向光性形成。     

低磷胁迫下水稻根系的发生及生长素的响应

 研究了5个磷浓度下（0,10,50,100,300 μmol/L）水稻植株的生物量以及水稻根系发生和伸长,并测定正常供P（300 μmol/L）与低磷(10 μmol/L)条件下水稻不同部位生长素浓度以及生长素外流蛋白OsPIN家族基因的表达情况。结果表明,与正常供P处理相比,随着供P浓度的降低,水稻地上部的干质量降幅显著,进而导致根冠比显著增加;与正常供P处理相比,低P处理的水稻根冠比增幅约为100%。水稻种子根、不定根和侧根的长度随供P浓度降低而显著增加,而不定根数及侧根密度随着供P浓度的降低而降低。与正常供P处理相比,低P处理的水稻倒1叶、根茎结合处和根系的生长素浓度显著上升,增幅分别为85%、161%和86%,差异达显著水平。RT PCR结果表明,与正常供P相比,低P处理24 h和96 h的水稻根系OsPIN5a表达上调。低P胁迫下水稻生长素合成和从地上部到根系极性运输的增强是水稻根系发生对低P胁迫响应的重要生理机制之一。     

Effect of Indoleacetic acid (IAA) on the Negative Phototropism of Rice Root

To explore the effects of IAA on negative phototropism of rice ( Oryza sativa L.) root, agar block containing IAA was unilaterally applied on root tip to examine the phototropic response of root to exogenous IAA, and microstructure of the bending part was observed with an optical microscope. The growth of seminal roots could be regulated by exogenous IAA as well as light,as a result the root bent towards the site treated, causing asymmetric growth of the root cells at the elongation zone and consequently bending growth. IAA concentration in the shaded side of adventitious root increased much greater at 1.5 h after the start of irradiation. The unequal lateral IAA distribution can be concluded to be the main cause for negative phototropism of rice root.     

生长素在水稻根负向光性反应中的作用

为了研究生长素在水稻根负向光性反应中的作用,用含有IAA的琼脂块贴附在种子根的根尖表面观测IAA对根的生长效应,用ELISA法测定根尖的IAA含量,并对种子根弯曲部分进行半薄切片,结果表明：(1) 种子根的生长方向既受光的调控也受外施的IAA的调控,根尖向贴有含IAA琼脂块的一侧弯曲生长;(2) 不定根在光照1.5 h后,背光侧的IAA含量明显大于向光侧;(3) 种子根尖发生弯曲生长是伸长区的细胞不均等生长所致。证实光引起根尖两侧IAA含量的差异导致根负向光性的形成。     

外源一氧化氮对盐胁迫下水稻根部脂质过氧化的缓解作用

 在100 mmol/L NaCl的胁迫条件下，研究了两种浓度的一氧化氮供体硝普钠（sodium nitroprusside，SNP）处理对水稻幼苗根部脂质过氧化的影响。低浓度（10 μmol/L）的SNP处理明显缓解盐胁迫下水稻幼苗根组织膜脂过氧化产物MDA的累积，显著提高可溶性蛋白含量，并诱导根系APX、POD和SOD活性的上升；而高浓度（50 μmol/L）SNP处理的效果则基本相反。表明外源低浓度NO可以通过提高盐胁迫下水稻幼苗根组织的抗氧化能力来缓解氧化损伤。     

Quantitative Trait Loci for Mercury Tolerance in Rice Seedlings

Mercury (Hg) is one of the most toxic heavy metals to living organisms and its conspicuous effect is the inhibition of root growth.However,little is known about the molecular genetic basis for root growth under excess Hg2+ stress.To map quantitative trait loci (QTLs) in rice for Hg2+ tolerance,a population of 120 recombinant inbred lines derived from a cross between two japonica cultivars Yuefu and IRAT109 was grown in 0.5 mmol/L CaCl2 solution.Relative root length (RRL),percentage of the seminal root length in +HgCl2 to-HgCl2,was used for assessing Hg2+ tolerance.In a dose-response experiment,Yuefu had a higher RRL than IRAT109 and showed the most significant difference at the Hg2+ concentration of 1.5 μmol/L.Three putative QTLs for RRL were detected on chromosomes 1,2 and 5,and totally explained about 35.7% of the phenotypic variance in Hg2+ tolerance.The identified QTLs for RRL might be useful for improving Hg2+ tolerance of rice by molecular marker-assisted selection.     

水稻根质膜Ca2+/H+反向转运体的存在及其特性

 由Ca2+/H+反向转运体蛋白的特异多肽制备的抗体，通过免疫印迹显示水稻根质膜上存在分子量为44.5 kD的蛋白。同位素闪烁记数法检测水稻根质膜上45Ca2+转运活性显示存在依赖于跨膜质子梯度的Ca2+/H+反向转运体，Mn2+可部分地抑制其转运活性。动力学分析表明，OsCAX2的Vmax=34.72 nmol/（min·mg）， Km=3.83 μmol/L，其最适pH为7.2。