水稻叶片对镉胁迫响应的蛋白质差异表达

为揭示水稻镉抗性的分子机理,以抗镉水稻品种P1312777和镉敏感水稻品种IR24为材料,在镉离子浓度为0(对照)、50和100 μmol·L-1条件下水培处理7 d,应用蛋白质组学方法分析了2种水稻叶片对镉胁迫响应的蛋白质差异表达.结果表明:镉胁迫下水稻PI312777叶片中共检测到差异表达蛋白质点31个,通过MALDI-TOF/MS分析,鉴定了其中的24个蛋白质(包括20个不同蛋白质,4个重复检出蛋白质);IR24叶片中共检测到差异表达蛋白质点19个,其中15个蛋白质得到鉴定.PI312777叶片鉴定出的20个蛋白质覆盖了IR24叶片鉴定的15个蛋白质,前者有4个与光合作用相关,11个与细胞防御代谢相关,3个与其他代谢相关,2个为功能未知蛋白.与对照相比,不同浓度镉胁迫下,抗镉水稻PI312777叶片中热激蛋白、谷胱甘肽还原酶、蛋白酶体α亚基6型、果糖1,6-二磷酸醛缩酶、硫氧还蛋白和DNA重组修复蛋白均上调表达;镉敏感水稻IR24叶片中热激蛋白、谷胱甘肽还原酶、蛋白酶体α亚基6型的表达无显著差异,果糖1,6-二磷酸醛缩酶和硫氧还蛋白则下调表达.此外,DNA重组修复蛋白仅在镉胁迫的PI312777叶片中表达.水稻PI312777比IR24具有更强的镉抗性与这些差异表达的蛋白质密切相关.     

不同耐性水稻幼苗根系对镉胁迫的形态及生理响应

采用水培试验,以两个耐镉性不同的水稻品种为材料,研究了不同浓度镉胁迫对水稻幼苗根系形态、根系活力、游离脯氨酸含量及抗氧化酶活性的影响。结果表明:低于5 μmol/L Cd胁迫对2个水稻品种总根长、根表面积、根体积、根干重、根系活力无明显影响,在1 μmol/L Cd时,甚至起促进作用。随Cd浓度增加表现出一定的抑制效应,秀水63在10 μmol/L Cd胁迫下根系形态、根系活力明显受到抑制,而秀水09在25 μmol/L Cd胁迫下明显受抑。随Cd胁迫浓度的增加,游离脯氨酸含量、丙二醛(MDA)含量、超氧化物歧化酶(SOD)、过氧化物酶(POD)活性均呈上升趋势,两品种相比,秀水09的游离脯氨酸含量、SOD和POD增幅大于秀水63,而MDA含量增幅小于秀水63;过氧化氢酶(CAT)活性变化表现为先上升后下降,10-100 μmol/L Cd胁迫下秀水63根系中CAT活性明显低于秀水09。总之,水稻对Cd毒害响应存在明显的品种差异,且Cd胁迫下根系生理响应的差异是品种间耐性差异的重要原因之一。     

热带地区不同水稻品种对土壤镉胁迫的生理生化响应

以热带地区主栽品种博优225和矮糯为材料,采用盆栽试验研究了不同水稻品种对土壤外源重金属Cd胁迫的耐性机理。结果表明:在各浓度镉胁迫下,2个品种水稻都是根累积的镉含量要高于茎叶和稻壳、糙米,即根>茎叶>稻壳>糙米;随着镉胁迫浓度的增加,细胞膜受损的伤害程度越大,矮糯质膜透性略高于博优225;随着Cd浓度的升高,水稻叶片丙二醛(MDA)、脯氨酸含量逐渐增加,且博优225的增加幅度明显大于矮糯,超氧化物歧化酶(SOD)和过氧化物酶(POD)活性均呈上升趋势,且博优225明显高于矮糯;过氧化氢酶(CAT)活性呈下降趋势;表明同样条件下博优225受Cd毒害更严重。     

磷高效水稻根系对低磷胁迫响应的差异蛋白分析

以IR71331为材料,通过水培试验,采用双向电泳分离不同磷浓度下（低磷浓度为0.5 mg·L^-1,对照为10 mg·L^-1）水稻生长3 d和6 d根系差异蛋白.结果表明：与对照相比,低磷胁迫下共有29个蛋白,其中3 d时间点有17个蛋白上调、11个下调、1个新增,6 d时间点有8个上调、19个下调、1个抑制表达、1个无明显变化 .经鉴定,其中的10个差异表达蛋白可归为信号转导相关蛋白、基因表达相关蛋白、代谢相关蛋白离子转运相关蛋白4个功能类群.信号转导相关蛋白分别为富含甘氨酸RNA结合蛋白和类似参与磷酸盐饥饿反应调控子;基因表达相关蛋白分别为推定的mRNA前体剪接因子SF2和推定的AAA蛋白酶家族FtsH;代谢相关蛋白分别为腺苷酸琥珀酸裂解酶、丝氨酸蛋白酶抑制剂(serpin)、S-腺苷蛋氨酸合成酶（SAM）和类似MYB类转录因子;离子转运相关蛋白分别为阳离子转运ATP酶和肌浆网膜蛋白.这些蛋白分别参与了信号识别、信号调控、mRNA 的剪接、信号传递、蛋白质降解、细胞体内离子转运和平衡等生理过程.其中serpin、SAM和MYB类转录因子是水稻响应低磷胁迫的关键蛋白.水稻根系对低磷胁迫存在着一个复杂的抗逆信号应答和代谢调控网络,其作用机理可以通过差异表达的蛋白质得以体现.     

镉胁迫下水稻OsPT1的表达及功能分析

OsPT1编码的水稻磷酸盐（Pi）转运蛋白在水稻生长发育、非生物胁迫应答等方面发挥重要的调控作用。前期研究表明OsPT1为镉（Cd）响应基因,但其在Cd胁迫下的功能及作用机制仍然未知。阐明OsPT1在Cd胁迫下的作用,并为低Cd水稻品种的选育奠定基础。通过生物信息学方法对该基因的序列特征、结构和功能进行分析和预测,利用实时荧光定量PCR（RT-qPCR）方法检测Cd胁迫下水稻不同组织、不同时间点OsPT1的相对表达量。此外,利用PCR的方法克隆OsPT1的编码序列,构建pGADT7-OsPT1重组质粒载体,并将其转入Δycf1 BY4741酵母菌株（Cd敏感酵母菌株）用以验证OsPT1对酵母Cd耐受性的影响。结果表明,OsPT1编码序列全长为1 584 bp,编码分子量为57.46 kD,由527个氨基酸构成的蛋白。在水稻基因组中该基因上游启动子区含有与光、厌氧、茉莉酸甲酯等环境和激素响应相关的调控元件。系统进化分析表明,水稻OsPT1与高粱SbPT1亲缘关系最近。基因的镉响应表达分析结果表明,与对照相比,经100 μmol/L Cd处理的水稻在1、6和12 h后,地上部分OsPT1的转录水平分别上调1.31、1.34和2.46倍;水稻根部OsPT1在处理1和6 h后分别上调1.28和1.14倍,但在Cd处理12 h后,其表达水平下调至处理前的0.62倍。转基因酵母Cd耐受性结果表明,与对照（0 μmol/L Cd）相比,经25 μmol/L Cd处理后,转OsPT1的酵母对Cd的耐受性有一定的下降。OsPT1可能在水稻应对Cd胁迫过程中发挥一定的作用。     

镉胁迫下小麦根系分泌物的变化

通过水培和砂培两种方法 ,研究了镉胁迫下小麦 ( Triticum aestivm L.)根系分泌物变化。通过研究镉对小麦根系分泌氨基酸和糖 ,根系分泌其它一些次生代谢物质等的影响 ,以及根系分泌物对蔬菜种子萌发的影响 ,探讨镉胁迫下植物根系的生理生态效应。研究结果表明 :镉胁迫下 ,小麦根系分泌物无论在量上还是质上都有变化。镉胁迫下根系分泌的电解质、糖类、氨基酸以及其它一些次生代谢物质都有所变化 ,但变化情况有差异。电解质外渗率、糖类随 Cd2 + 浓度升高而增加 ,在低浓度 Cd2 + 作用下 ( 0 .5 mg/ L) ,随处理浓度的升高 ,氨基酸分泌量增加 ;当处理浓度高于相应浓度时 ,氨基酸分泌量随浓度升高而减少。随 Cd2 + 浓度升高 ,次生代谢物分泌种类减少。这说明小麦通过改变根分泌作用而缓解镉危害。由于根系分泌次生代谢物的活动比较复杂 ,因而根系的分泌作用有待于进一步研究。根系分泌物中可能含有某些物质抑制萝卜、白菜种子萌发 ,这是涉及到植物他感作用的问题 ,值得并有待于进一步研究。初步分析小麦根系在不同的环境中对镉胁迫的响应方式 ,为以后根分泌物的深入研究提供了一定的思路 ,也为镉污染区农作物的合理栽培提供新的参考资料。     

镉胁迫下杞柳对金属元素的吸收及其根系形态构型特征

采用水培方法,研究了杞柳(Salix integra)2个品种在0-90 μmol/L Cd处理下不同组织对Cd的吸收和积累规律,探讨了镉胁迫下杞柳根系形态学功能响应特征以及对矿质离子吸收的影响。结果发现,镉在杞柳2个品种不同组织的含量均表现为根>韧皮部>木质部>叶。2个品种地上部组织对镉的吸收和积累规律相似:在0-70 μmol/L Cd处理浓度范围内,随着溶液Cd浓度的增加,叶、木质部、韧皮部中镉的含量逐渐增加,到50 μmol/L时,镉在地上部组织的含量达到最高,当Cd处理浓度达90 μmol/L时,地上部各组织中Cd含量出现下降趋势。而2个品种根系对Cd的吸收则不同,"微山湖"品种在溶液Cd达70 μmol/L时,根系镉的积累量最大,在90 μmol/L时,根系Cd的积累量明显下降;"一枝笔"品种根系在0-90 μmol/L范围内,根系Cd的积累量均为增加趋势。通过分析2个品种根系形态学参数变化发现,Cd胁迫抑制了杞柳2个品种根的伸长,促进了"微山湖"品种根的径向生长,导致根系平均直径增加;进一步对叶片矿质营养状态分析发现, Ca、Mn的吸收受镉胁迫的影响较大,在高浓度镉胁迫下(50-70 μmol/L)"微山湖"和"一枝笔"对Ca和Mn的吸收明显下降;Cd胁迫对Fe在叶片的含量影响不明显;同时发现,在50 μmol/L Cd处理下,Cu²⁺在叶片的积累明显增加。由此可见,杞柳2个品种间对镉的吸收和积累差异主要体现在根系,地上部对镉毒害的响应差异不大。     

铜绿假单胞菌对镉胁迫苗期水稻根系活力及叶片生理特性的影响

为揭示耐镉铜绿假单胞菌缓解镉胁迫水稻的生理效应,以无镉处理为对照,通过添加菌液、空载体、菌剂及20μmol·L^-1 Cd进行水培试验,分析了菌株对苗期水稻根系活力及叶片生理特性的影响.结果表明:镉胁迫显著抑制了水稻的根系活力,降低了叶片光合效率、抗氧化酶活性及可溶性蛋白、类黄酮与总酚含量,提高了叶片丙二醛(MDA)和超氧阴离子(O2-)含量.与镉处理相比,添加菌液、菌剂处理的水稻根系活力分别提高了36.1%~42.5%、49.4%~53.0%;叶片净光合速率提高了118.5%~147.1%、137.6%~156.9%;可溶性蛋白含量提高了37.0%~49.3%、37.7%~72.6%.菌剂处理的水稻叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性分别比Cd处理提高了36.9%~42.6%、82.7%~92.6%、43.3%~52.2%,菌液处理的SOD、POD、CAT则分别比Cd处理提高了25.8%~36.6%、40.9%~55.9%、24.0%~29.2%,菌剂对水稻叶片抗氧化酶的促进效应显著高于菌液;菌剂、菌液处理的水稻叶片MDA含量分别比Cd处理降低了44.8%~54.7%、29.4%~41.9%;O2-含量减少了9.9%~10.2%、3.0%~7.1%;菌剂处理后类黄酮、总酚含量分别比Cd处理提高了125.4%~135.7%、100.8%~119.4%;菌液处理后则分别提高了139.4%~146.7%、115.0%~134.7%.可见,铜绿假单胞菌及其菌剂通过提高苗期水稻根系活力、光合作用促进了苗期水稻的生长.铜绿假单胞菌通过增强水稻抗氧化酶活性、提高类黄酮和总酚等抗氧化物质含量,表现出显著的缓解镉胁迫效应.     

镉腔迫下小麦根系分泌物的变化

通过水培和砂培两种方法,研究了镉胁迫下小麦（Triticum aestivm L.)根系分泌物变化。通过研究镉对小麦根系分泌氨基酸和糖,根系分泌其它一次次生代谢物质等的影响,以及根系分泌物对蔬菜种子萌发的影响,探讨镉胁迫下植物根系的生理生态效应。研究结果表明：镉胁迫下,小麦根系分泌物无论在量上还是质上都有变化。镉胁迫下根系分泌的电解质,糖类,氨基酸以及其它一些次生代谢物质都有所变化,但变化情况有差异。电解质外渗率,糖类随Cd^2＋浓度升高而增加,在低浓度Cd^2 作用下（0.5mg/L),随处理浓度的升高,氨基酸分泌量增加,当处理浓度高于相应浓度时,氨基酸分泌量随浓度升高而减少。随Cd^2 浓度升高,次生代谢物分泌种类减少。这说明小麦通过改变根分泌作用而缓解镉危害。由于根系分泌次生代谢物的活动比较复杂,因而根系的分泌作用有待于进一步研究。根系分泌物中可能含有某些物质抑制萝卜,白菜种子萌发,这是涉及到植物他感作用的问题,值得并有待于进一步研究。初步分析小麦根系在不同的环境中对镉胁迫的响应方式,为以后根分泌物的深入研究提供了一定的思路,也为镉污染区农作物的合理栽培提供新的参考资料。     

镉胁迫对芥蓝根系质膜过氧化及ATPase活性的影响

水培条件下,以“香港白花”芥蓝品种为供试材料,研究4种不同浓度镉（0、1.0、2.0、4.0、8.0 mg/L Cd）处理对芥蓝幼苗根系质膜过氧化及ATPase活性的影响。结果表明,与对照相比,随着Cd处理浓度的增加,芥蓝根系活力呈现降低的变化趋势,而根系超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性以及丙二醛（MDA）、H2O2含量和O2-产生速率表现出升高的趋势,表明芥蓝受到活性氧物质的胁迫。1.0、2.0 mg/L Cd浓度处理下的H2O2含量与对照差异不显著,而O2-产生速率则在1.0 mg/L浓度处理下与对照差异不显著。随着Cd处理浓度的增加,芥蓝根系质膜H -ATPase和Ca2 -ATPase活性呈现出先增后减的变化趋势。1.0 mg/LCd浓度处理时,H -ATPase和Ca2 -ATPase活性与对照差异不显著（P > 0.05）,而在2.0、4.0和8.0 mg/LCd处理时,两种ATPase活性显著降低（P < 0.05）,并且与膜脂过氧化水平呈极显著的负相关（R2 > 0.969）。因此,低浓度Cd处理对芥蓝根系质膜两种ATPase活性影响较小,较高浓度Cd处理使芥蓝根系活力和质膜ATPase的损伤加重。     

Glutathione reduces the inhibition of rice seedling root growth caused by cadmium

We investigated the effects of agents known to affect cellular glutathione (reduced form, GSH) levels on the growth of rice seedlings treated with Cd. CdCl₂ was more effective than CdSO₄ in inhibiting root growth. However, CdCl₂ had no effect on shoot growth. GSH, a substrate for phytochelatin synthesis, was effective in counteracting growth inhibition of roots by CdCl₂. Root growth in the CdCl₂ medium was found also to be enhanced by the addition of L-glutamic acid and L-cysteine, both of which are substrates for GSH formation. Buthionine sulfoximine, an inhibitor of GSH synthesis, rendered the roots susceptible to growth inhibition by Cd. Our results suggest that GSH level may play a role in regulating Cd-inhibited growth of rice roots.Abbreviations BSO buthionine sulfoximine - GSH reduced form glutathione     

Growth direction of nodal roots in rice: its variation and contribution to root system formation

The root system of a rice plant (Oryza sativa L.) consists of numerous nodal roots and their laterals. The growth direction of these nodal roots affects the spatial distribution of the root system in soil, which seems to relate to yield and lodging resistance. The growth angle of a nodal root varies with the type and timing of emergence of the nodal root. The body of a rice plant can be recognized as an integrated set of shoot units, each unit consisting of an internode with a leaf and several roots. Nodal roots formed at the apical part of a shoot unit often elongate horizontally, whereas those formed at the basal part of the shoot unit show various growth directions depending on both the growth stages of the plant and the environmental conditions. Moreover, nodal roots that emerge from the most basal shoot unit of a tiller are usually thick and grow downwards. External factors such as planting density and nitrogen application affect the growth direction of nodal roots, probably partly because of the changing tillering pattern of the shoot. In addition to the growth angle of nodal roots, size of nodal roots may be another important factor determining the spatial distribution of the root system in soil.     

Plant regeneration from protoplasts of indica rice: genotypic differences in culture response

Summary Fourteen varieties of indica rice (Oryza saliva L.) were examined for their capacity for plant regeneration from protoplasts using the nurse culture methods developed for japonica rice. Calli induced from germinating seeds were grouped into two types: type I, white and compact; type II, yellow and friable. In four varieties producing type II callus, colony formation (2%–4.5%) and plant regeneration (2%–35%) were observed. The inability to develop suspension cultures was a major obstacle in regenerating plants from protoplasts of the remaining rice varieties studied.     

Disturbed ammonium assimilation is associated with growth inhibition of roots in rice seedlings caused by NaCl

The effects of NaCl on changes in ammonium level and enzyme activities of ammonium assimilation in roots growth of rice (Oryza sativa L.) seedlings were investigated. NaCl was effective in inhibiting root growth and stimulated the accumulation of ammonium in roots. Accumulation of ammonium in roots preceded inhibition of root growth caused by NaCl. Both effects caused by NaCl are reversible. Exogenous ammonium chloride and methionine sulfoximine (MSO), which caused ammonium accumulation in roots, inhibited root growth of rice seedlings. NaCl decreased glutamine synthetase and glutamate synthase activities in roots, but increased glutamate dehydrogenase activity. The growth inhibition of roots by NaCl or MSO could be reversed by the addition of L-glutamic acid or L-glutamine. The current results suggest that disturbance of ammonium assimilation in roots may be involved in regulating root growth reduction caused by NaCl.Abbreviations GDH glutamate dehydrogenase - GOGAT glutamate synthase - GS glutamine synthetase - MSO methionine sulfoximine     

Cd induced changes in proline level and peroxidase activity in roots of rice seedlings

The effects of Cd on changes in proline level and peroxidase activity in roots of rice seedlings were investigated. CdCl₂ was effective in inhibiting root growth and in accumulating proline in roots. The inhibition of root growth by Cd is reversible. The reduction of root growth induced by Cd is closely associated with accumulation of proline in roots. External application of proline markedly inhibited root growth of rice seedlings in the absence of Cd. Ionically bound, but not soluble, peroxidase activity in roots was increased by CdCl₂. Proline treatment also resulted in an increase in ionically bound peroxidase activity in roots. The relationship between growth inhibition of roots induced by Cd and changes of proline level and peroxidase activity is discussed.Abbreviations POX peroxidase     

Ammonium nutrition increases photosynthesis rate under water stress at early development stage of rice (Oryza sativa L.)

An hydroponic experiment with a simulated water stress induced by PEG (6000) was conducted in a greenhouse to study the effects of nitrate (NO₃ ⁻), ammonium (NH₄ ⁺) and the mixture of NO₃ ⁻ and NH₄ ⁺, on water stress tolerance of rice seedlings. Rice (Shanyou 63) was grown under non- or simulated water stress condition (10% (w/v) PEG, MW6000) with the 3 different N forms during 4 weeks. Under non-stressed condition no difference was observed among the N treatments. Under simulated water stress, seedlings grown on N-NO₃ ⁻ were stunted. Addition of PEG did not affect rice seedling growth in the treatment of only NH₄ ⁺ supply but slightly inhibited the rice seedling growth in the treatment of mixed supply of NO₃ ⁻ and NH₄ ⁺. Simulated water stress, when only N-NH₄ ⁺ was present, did not affect leaf area and photosynthesis rate, however, both parameters decreased significantly in the NO₃ ⁻ containing solutions. Under water stress, Rubisco content in newly expanded leaves significantly increased in the sole NH₄ ⁺ supplied plants as compared to that in plants of the other two N treatments. Under water stress, the ratio of carboxylation efficiency to Rubisco content was, respectively, decreased by 13 and 23% in NH₄ ⁺ and NO₃ ⁻ treatments, respectively. It is concluded that, water stress influenced the Rubisco activity than stomatal limitation, and this effects could be regulated by N forms. Responsible Editor: Herbert Johannes Kronzucker. Shiwei Guo and Gui Chen contributed equally to this paper.     

Sink anatomy in relation to solute movement in rice (Oryza sativa L.): A summary of findings

The results of studies on assimilate and water transport in the developing caryopsis of rice are summarised. Evidence is presented for a symplastic movement of solutes as far as the aleurone layer. However, transport into the apoplast at the nucellus/aleurone interface appears to be a necessary step due to the absence of plasmodesmata at this site. It is suggested that water leaves the caryopsis during grain filling by the isolated cell walls of the pigment strand, the suberised walls of these cells functioning to isolate the apoplast from the symplast and thereby allowing opposing fluxes of water and assimilates to occur in the dorsal region of the grain.     

Using iron fertilizer to control Cd accumulation in rice plants: A new promising technology

Effects of two kinds of iron fertilizer, FeSO4 and EDTA·Na2Fe were studied on cadmium accumulation in rice plants with two rice genotypes, Zhongzao 22 and Zhongjiazao 02, with soil culture systems. The results showed that application of iron fertilizers could hardly make adverse effects on plant growth and rice grain yield. Soil application of EDTA·Na2Fe significantly reduced the Cd accumulation in rice roots, shoots and rice grain. Cd concentration in white rice of both rice genotypes in the treatment of soil application of EDTA·Na2Fe was much lower than 0.2 mg/kg, the maximal Cd permission concentra- tion in cereal crop foods in State standard. However, soil application of FeSO4 or foliar application of FeSO4 or EDTA·Na2Fe resulted in the significant increase of Cd accumulation in rice plants including rice grain compared with the control. The results also showed iron fertilizers increased the concentra- tion of iron, copper and manganese element in rice grain and also affected zinc concentration in plants. It may be a new promising way to regulate Cd accumulation in rice grain in rice production through soil application of EDTA·Na2Fe fertilizers to maintain higher content of available iron and ferrous iron in soils.     

Growth and CO2 assimilation of lowland rice in response to timing and method of N fertilization

In order to understand more clearly the dynamics of rice (Oryza sativa L.) yield development in relation to N fertilization, a field experiment was conducted in Laguna, Philippines. The soil, a Maahas clay (Aquic Hapludalf), was flooded, puddled, and then planted with 20-day-old seedlings of IR64 rice. Treatments were six combinations of N fertilizer timing and method: (1) 0 N control; (2) prilled urea broadcast at 15 and 60 days after transplanting (DAT) (BR-LATE); (3) prilled urea injected with a spring auger applicator at 15 DAT and broadcast at 60 DAT (INJ-LATE); (4) prilled urea broadcast and incorporated at 0 DAT and broadcast at 40 DAT (BR-EARLY); (5) urea super granules (USG) manually deep-placed at 3 DAT and prilled urea broadcast at 40 DAT (DP-EARLY); and (6) USG manually deep-placed at 3 DAT (DP). Except for the control, all treatments received a total of 58 kg N ha^-1. Yield results were consistent with those of other experiments, namely, DP had the highest yields, the early-split treatments (BR-EARLY and DP-EARLY) were second best, followed by the late-split treatments (BR-LATE and INJ-LATE), with the control having the lowest yield. Sequential harvest results showed that the advantages of DP in terms of dry matter assimilation, tillering, and leaf area index (LAI) were expressed later in the season. For all treatments, midday net CO₂ assimilation (Ac) peaked around 48 DAT, approximately panicle initiation. Grain yield was highly correlated with midday Ac at panicle initiation and flowering but not at other growth periods. Rates of midday Ac and night respiration depended strongly on shoot N content. We conclude that N application method and timing should be designed to give high shoot N content at panicle initiation and flowering, and that DP satisfied this requirement best among the treatments tested.