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.     

低磷胁迫水稻根部基因表达谱研究

磷是植物体内重要的营养元素．土壤中含磷总量丰富,但能被植物直接吸收利用的可溶性磷含量却很低,这成为制约农作物产量的重要因素．本研究利用水稻寡核苷酸芯片分析了水稻根部在正常营养条件和低磷胁迫6,24,72h3个时间点的全基因组表达谱．和正常营养条件下相比,低磷胁迫水稻根部共发现795个差异表达基因．差异表达基因功能分析发现：（1）磷酸盐转运蛋白、酸性磷酸酶、RNA酶等基因上升表达;（2）糖酵解等与能量代谢相关基因先上升后下降表达;（3）氮吸收和脂代谢相关基因改变其表达;（4）蛋白质降解、细胞衰老相关基因上升表达;（5）部分跨膜转运蛋白基因表达上调．研究结果为进一步揭示植物低磷胁迫反应机制,改善作物对磷吸收利用效率提供了有用的信息．     

高产磷高效水稻磷素吸收利用特征

通过正常供磷的大田试验（2011年）,以产量和磷籽粒生产效率为指标,将27份中熟水稻亲本材料划分为4个类型,再通过正常和低磷处理的土培试验（2012年）,筛选出高产磷高效水稻材料,并探讨各种磷效率对产量的贡献率.结果表明： 结合两年的试验结果,供试材料的产量和磷利用效率均存在显著的基因型差异,筛选出GR泸17/矮TTP//泸17_2（QR20）为高产磷高效材料.在两个供磷水平下,QR20的产量和磷利用效率均显著高于低产磷低效材料玉香B,其产量分别是玉香B的1.96和1.92倍.大田和土培试验结果均表明,磷积累量对产量的贡献率均高于磷籽粒生产效率和磷收获指数.正常供磷条件下,磷积累量和磷籽粒生产效率对产量的贡献率差异不大,低磷条件下差异较大（66.5%和26.6%）,磷收获指数对产量的贡献率最低,最高仅为11.8%（土培）.土培试验中,正常供磷条件下,拔节-抽穗阶段的磷积累量对产量和磷收获指数的贡献率最高,分别为93.4%和85.7%,对磷籽粒生产效率的贡献率为41.8%;在低磷条件下,分蘖-拔节阶段的磷积累量对产量和磷籽粒生产效率的贡献率最高,分别为56.9%和20.1%,对磷收获指数的贡献率为16.0%.土培正常供磷条件下,水稻QR20的产量、磷积累量和磷收获指数相对于低磷处理分别增加了20.6%、18.1%和18.2%,差异显著.综上,磷效率对水稻产量的贡献率大小依次为磷吸收效率＞利用效率＞转运效率;正常供磷条件下,拔节-抽穗阶段的磷积累量对产量的贡献率最高,低磷胁迫下,分蘖-拔节阶段的磷积累量对产量的贡献率最高,这两个阶段可能是水稻高产磷高效协调统一的关键时期.
     

低磷胁迫下甘蓝型油菜酸性磷酸酶对磷效率的贡献分析

为探讨低磷胁迫下甘蓝型油菜酸性磷酸酶活性的基因型差异及其与磷效率的关系, 采用土培实验研究了磷高效基因型102和磷低效基因型105对有机磷和无机磷的利用及其根际土壤酸性磷酸酶活性差异; 并采用水培实验研究了甘蓝型油菜根系分泌的酸性磷酸酶及不同叶片酸性磷酸酶的活性差异. 结果表明, 低磷胁迫能诱导根系及根系分泌的酸性磷酸酶活性升高; 土培条件下, 由于酸性磷酸酶的有效性受较多因素影响, 植物的磷营养和磷吸收效率与根系分泌的酸性磷酸酶活性并不直接相关; 缺磷胁迫下重组自交系群体叶片酸性磷酸酶活性与磷利用效率呈显著正相关, 进一步表明低磷诱导的植株叶片酸性磷酸酶活性升高能促进体内磷的再利用, 从而提高磷利用效率.     

耐低磷水稻基因型筛选指标的研究

采用溶液培养试验,并结合大田试验,研究和探讨了耐低磷水稻基因型的筛选指标.结果表明,溶液培养试验中,在正常供磷和低磷胁迫条件下,在所有调查性状中水稻单株干重都具有较大的基因型间变异(CV分别为21.73%和19.54%).在所有调查性状的相对值中,相对单株干重(低磷胁迫/正常供磷)也具有较大的基因型间变异(CV为19.60%);相关分析表明,相对单株干重与相对根干重、相对株高、相对单株吸磷量、相对地上部磷积累、相对磷利用效率和相对植株磷浓度均呈极显著正相关(P＜0.01).因此,水稻相对单株干重可以作为苗期筛选水稻耐低磷基因型的一个筛选指标.溶液培养试验中水稻的相对单株干重和大田试验中水稻的相对稻谷产量(不施磷/施磷)没有显著相关性,因此溶液培养试验的相对单株干重不能作为评价大田试验中水稻耐低磷能力的指标.低磷营养液培养的水稻体内磷利用效率与缺磷土壤生长的水稻体内磷利用效率呈极显著正相关.因此,直接以低磷营养液培养水稻苗期体内磷利用效率作为筛选指标,然后进行大田试验验证,是一条筛选水稻磷高效利用基因型的有效途径.     

缺血／再灌注损伤对心肌细胞核膜NTPase活性和mRNA出核转运的影响

目的：观察家兔心肌缺血／再灌注（I／R）损伤对心肌细胞核膜核苷三磷酸酶（NTPase）动力学及mRNA出核转运的变化。方法：制备家兔离体心脏I／R模型,密度梯度离心法制备心肌细胞核膜,按Tiffany等的方法测定NTPase活性及mRNA的转运率。结果：心肌细胞核膜NTPase在以ATP或GTP为底物时,与对照组比较,I／R组最大反应速率（Vmax)分别降低26％及22％（P＜0．01）。Km值分别升高54％及72％（P＜0．01）;而缺血组Vmax及Km值较对照组无明显差异。以ATP或GTP启动反应时I／R组10min内心肌细胞核mRNA转出率分别较对照组降低27％及32％（P＜0．01）;而缺血组与对照组相比并无显著性差异（P＞0．05）。结论：心肌缺血／再灌注损伤时,心肌细胞核膜受损,核膜上NTPase活性降低,导致细胞核膜mRNA出核受阻,从而可能影响蛋白质的合成表达。     

南方水稻氮素吸收与利用效率的基因型差异及评价

 以南方籼型水稻（Oryza sativa）品种为试验材料进行大田试验,以探讨提高水稻氮素吸收与利用效率的基因型潜力。结果表明,除早季分蘖期氮素积累量、干物质生产效率和抽穗期氮素积累量以及晚季氮素运转效率外,各基因型氮素吸收与利用效率存在显著或极显著的差异,提高水稻氮素吸收与利用效率的基因型潜力很大。基因型生育期对其氮素吸收与利用效率产生重要影响,生育期较长的基因型其氮素吸收效率、稻谷和干物质生产效率以及农艺效率较高。杂交稻氮素的生产效率、农艺效率、回收效率和收获指数较常规稻高,但二系杂交稻并没有比三系杂交稻明显提高。通过排序方式对各基因型氮素吸收与利用效率进行评价的结果表明,不同氮素吸收与利用效率指标的排序以及同一指标早晚季的排序均存在较大差异。氮素吸收与利用效率经标准化后的综合排序可对各基因型的氮素吸收与利用效率进行综合评价,吻合系数则可较好地反映各基因型早晚季氮素吸收与利用效率的排序。     

模拟氮沉降对低磷胁迫下马尾松不同家系根系分泌和磷效率的影响

近年来大气氮(N)沉降的增加, 导致森林土壤中有效N含量增加、N:P发生改变, 研究N沉降对低磷(P)胁迫下林木根系分泌和P效率的影响具有重要意义。该文以马尾松(Pinus massoniana)家系作为试验材料, 设置模拟N沉降与同质低P (介质表层与深层均缺P)、异质低P (介质表层P丰富、深层缺P)耦合的二年生盆栽实验, 系统研究了模拟N沉降对低P胁迫下马尾松根系分泌性酸性磷酸酶(APase)活性、有机酸分泌以及P效率的影响。结果表明: (1)同质低P和异质低P下, 模拟N沉降均显著提高了植株N:P化学计量比、增加了P素的相对匮乏程度, 从而诱导根系增加了APase和有机酸的分泌, 而同质低P比异质低P下增加幅度更大, 其中有机酸分泌均与马尾松生长呈正相关关系, 而APase活性与P效率相关性较小; (2)同质低P下, N沉降虽然增加了根系分泌, 但未提高马尾松P素吸收和生长量, 其原因在于, 同质低P下植株N:P过高, 因而植株对N沉降敏感性低; 在异质低P下, 植株表现为N、P共同限制, 因而对N敏感性较高, N沉降增加了根系分泌, 同时提高了N和P吸收效率、增加了生物量; (3)马尾松根系分泌对模拟N沉降的响应存在较大的家系差异。同质低P下, 家系71×20的有机酸分泌和生物量对N沉降的响应幅度较大; 异质低P下, 家系36×29、71×20和73×23对N沉降的响应幅度较大。     

水稻苗期磷高效基因型筛选研究

采用难溶性磷酸盐Ca3 (PO4) 2 为唯一磷源 ,在pH值为 5 .5条件下产生相对高浓度低磷胁迫及以NaH2 PO4为磷源配制P浓度为 0 .5mg·L-1的相对低浓度低磷胁迫的两个水培环境 ,分别对不同基因型水稻的磷效率进行评价 .以相对分蘖干重 (RTW )、相对总生物量 (RPW )、相对分蘖数 (RTN)、相对根系干重(RRW )、相对地上部干重 (RSW )、相对叶龄 (RLA)和相对株高 (RPH)作为耐性指标进行相关分析 .结果表明 ,供试材料的磷效率存在极显著差异 ,若以能产生分蘖的相对高浓度低磷胁迫进行筛选时 ,相对分蘖干重、相对地上部干重、相对总生物量可作为较好的筛选指标 ,其中相对分蘖干重不仅与其它指标间的相关性强 ,且品种间差异和变异系数均较大 ,能准确、灵敏地反映不同基因型间的耐低磷胁迫能力 ;若采用相对低浓度的低磷胁迫对不同基因型水稻进行耐低磷种质筛选时 ,筛选指标则不同 ,最好的单一筛选指标应是相对地上部干重或相对总生物量 .     

不同磷效率小麦品种的磷吸收特性

在丰磷、缺磷条件下,对不同磷效率小麦品种的磷吸收特性进行研究。缺磷条件下,不同磷效率品种成熟期的植株全磷量和生育中后期（挑旗-成熟期）植株磷累积量均以磷高效品种最高,中效品种次之,低效品种最低。不同磷效率品种拔节期、挑旗期和成熟期的磷利用效率差异较小。表明磷高效小麦品种在缺磷条件下子粒产量形成能力的提高。与生育中后期植株具有相对较强的磷素吸收能力有关。缺磷条件下,不同磷效率品种在生育中后期的根系TTC还原力和可溶蛋白含量也以高效品种最高,中效次之,低效最低。表明磷高效小麦品种植株生育中后期根系具有较强的生理功能,是其在缺磷务件下吸磷量增加、产量相对明显提高的重要生理基础。研究表明,不同磷效率小麦品种在磷胁迫条件下的根系酸性磷酸化酶（APase）活性存在显著差异,并在小麦磷吸收效率的调控中具有重要作用。     

应用AFLP与RFLP标记研究水稻磷吸收与利用率的数量性状位点

To identify genetic factors underlying phosphorus (P) uptake and use efficiency under low-P stress in rice (Oryza sativa L.), 84 selected genotypes (recombinant inbred lines) and their parents (which differed in tolerance for low-P stress) “IR20” and IR55178-3B-9-3, were cultured in liquid medium supplemented with adequate and low P levels in a greenhouse. Plants were sampled after 6 weeks in culture for measurements of plant dry weight, P concentration, P uptake and P use efficiency under both P sufficient and stress conditions. A total of 179 molecular markers, including 26 RFLPs and 153 AFLPs, mapped on all 12 chromosomes of rice based on the 84 selected genotypes were used to detect the quantitative trait loci (QTLs) underlying tolerance for low-P stress. Three QTLs were detected on chromosomes 6, 7 and 12, respectively, for relative plant dry weight (RPDW) and relative P uptake (RPUP). One of the QTLs flanked by RG9 and RG241 on chromosome 12 had a major effect which explained about 50% of the variations in the two parameters across the population. The results coincided with the QTLs for low-P stress based on relative tillering ability from the same population from a cross between Nipponbare and Kasalath under soil condition. The identical major QTL for P uptake and plant growth under low-P stress in both liquid medium and soil strongly suggests that the ability of P uptake mainly controls rice tolerance for low-P stress.     

How do plants achieve tolerance to phosphorus deficiency? Small causes with big effects

Genotypic differences in phosphorus (P) uptake from P-deficient soils may be due to higher root growth or higher external root efficiency (micrograms of P taken up per square centimeter of root surface area). Both factors are highly interrelated because any additional P provided by externally efficient roots will also stimulate root growth. It will be necessary to separate both factors to identify a primary mechanism to formulate hypotheses on pathways and genes causing genotypic differences in P uptake. For this purpose, a plant growth model was developed for rice (Oryza sativa) grown under highly P-deficient conditions. Model simulations showed that small changes in root growth-related parameters had big effects on P uptake. Increasing root fineness or the internal efficiency for root dry matter production (dry matter accumulated per unit P distributed to roots) by 22% was sufficient to increase P uptake by a factor of three. That same effect could be achieved by a 33% increase in external root efficiency. However, the direct effect of increasing external root efficiency accounted for little over 10% of the 3-fold increase in P uptake. The remaining 90% was due to enhanced root growth as a result of higher P uptake per unit root size. These results demonstrate that large genotypic differences in P uptake from a P-deficient soil can be caused by rather small changes in tolerance mechanisms. Such changes will be particularly difficult to detect for external efficiency because they are likely overshadowed by secondary root growth effects.     

Effect of fertilization on exudation,dehydrogenase activity,iron-reducing populations and Fe++ formation in the rhizosphere of rice (Oryza sativa L.) in relation to iron toxicity

Summary To explain the mechanism of iron toxicity, greenhouse and growth chamber (¹⁴CO₂ atmosphere) experiments were carried out. In pot experiments (with a typical iron-toxic soil and a fertile clay) we studied the effect of N, P, K and Ca+Mg fertilization (alone or in combination) on dehydrogenase activity, Fe⁺⁺ formation, and the populations of iron-reducing bacteria in the rhizosphere of rice IR22 and IR42. Fe uptake by the plants was measured at regular intervals. Dehydrogenase activity, the number of N₂-fixing iron-reducing bacteria, and the formation and uptake of Fe⁺⁺ decreased with increased supply of K, Ca, and Mg. This effect was clearer with IR22 (susceptible to iron toxicity) than with IR42 (releatively tolerant). Increased exudation and Fe uptake by IR36 at low nutrient and high Fe supply were recorded in a growth chamber experiment. Nutritional conditions, exudation rate (a measure of metabolic root leakage), the iron-reducing activity of the rhizosphere, and Fe⁺⁺ uptake by wetland rice appear to be clearly related. Iron toxicity is considered a physiological disorder caused by multiple nutritional soil stress rather than by a low pH and high Fe supply per sé.     

Study on the effect of hypergravity stress on L-glutamate uptake by rat brain nerve endings

Using rat brain synaptosomes, we have investigated the effect of hypergravity on the kinetic parameters of Na(+)-dependent, high-affinity L-glutamate transport activity. The time-course of L-[14C]-glutamate uptake and dependence of L-[14C]-glutamate uptake velocity on glutamate concentrations were analyzed. K(m) and Vmax of this process have been determined. The hypergravity stress was created by centrifugation of rats for 1 hour at 10 g. We observed no differences in K(m) values between the control rats (10.7 +/- 2.5 microM) and animals exposed to hypergravity (6.7 +/- 1.5 microM). The similarity of this parameter for the two studied groups of animals showed that affinity of glutamate transporter to substrate was not sensitive to hypergravity stress. In contrast, the maximal velocity of glutamate uptake changed in hypergravity conditions. Vmax reduced from 12.5 +/- +/- 3.2 nmol/min per 1 mg of protein (control group) to 5.6 +/- 0.9 nmol/min per 1 mg of protein (animals, exposed to hypergravity stress). The possible mechanisms of attenuation of the glutamate transporter activity without modifying K(m) of glutamate uptake were discussed.     

Aluminium effects on growth, nutrient net uptake and transport in 3 rice (Oryza sativa) cultivars with different sensitivity to aluminium

Varietal differences in net nutrient uptake rate and transport efficiency in the presence of aluminium have seldom been investigated in rice. Therefore, effects of Al on growth, uptake and transport of macronutrients (K, P, Ca, and Mg) and micronutrients (Fe, Zn, Cu, and Mn) were evaluated in 3 rice cultivars (BG35, DA14 and IR45) with different Al sensitivity. The plants were grown in nutrient solution at pH 4.1. An initial growth was completed in the time interval 1 to 5 days immediately before the addition of Al. The final growth period with Al (0, 140, 280 or 560 μ M ) was completed on day 26. With Al, a comparatively high P accumulation occurred in shoots and roots of the Al tolerant cultivar BG35. In contrast, the Al sensitive cultivar IR45 maintained a relatively high Ca accumulation during the Al treatment. A reduced total net uptake rate of P and Ca by IR45 in the time period 5 to 26 days was due to both a reduced root fresh weight and a reduced net uptake rate per g fresh weight of root. Moreover, net Ca transport to the shoots higher than net uptake rate in DA14 and IR45 at > 140 μ M Al during the test period suggests restricted Ca uptake by the roots in combination with a continuous net loss of Ca from the roots to the shoots as time proceeds. In the case of Mg and Mn, there was a general reduction of net uptake rates, irrespective of Al sensitivity of cultivars. With Al treatment, comparatively high accumulation of Fe, Zn and Cu occurred in the roots of IR45, concomitant with a high net Zn and Cu uptake rate. It is concluded that differences in Al sensitivity among rice cultivars BG35, DA14 and IR45 are not primarily linked to the depressed internal Mg or Mn status of the plants but rather to changes in the uptake and distribution of Ca and P.     

Differential changes in photosynthetic capacity, 77 K chlorophyll fluorescence and chloroplast ultrastructure between Zn-efficient and Zn-inefficient rice genotypes (Oryza sativa) under low zinc stress

The relationship of zinc (Zn) efficiency in rice to differential tolerance of photosynthetic capacity and chloroplast function to low Zn stress was studied using Zn-efficient (IR8192) and Zn-inefficient (Erjiufeng) rice genotypes ( Oryza sativa L.). Zinc deficiency caused extensive declines in leaf chlorophyll (Chl) content, ratios of chl a:b, Pn, Fv/Fm and Fv/Fo, indicating that the intrinsic quantum efficiency of the photosystem II (PSII) units was damaged. A greater decline was observed in the inefficient genotype (Erjiufeng) than the efficient genotype (IR8192). The 77 K chl fluorescence emission spectrum revealed that Zn deficiency blocked energy spillover from PSII to PSI and more excitation energy was distributed to PSII in IR8192 than Erjiufeng. The spectrum of Zn-deficient Erjiufeng was completely disordered, implying that the photosynthetic centers were seriously damaged. Electron microscopy showed that Zn deficiency caused a severe damage to the fine structure of chloroplasts, but IR8192 had a better preserved chloroplast ultrastructure as compared with Erjiufeng. These differences may result from the higher levels of the antioxidant enzyme activities and lower oxidant stress level in IR8192. These results indicate that Zn deficiency decreases leaf photosynthetic capacity primarily by reducing the number of PSII units per unit leaf area, and also reducing the photochemical capacity of the remaining PSII units. Therefore, the maintenance of more efficient photochemical capacity under low Zn stress is a key factor for the high Zn efficiency in rice, which may result from less antioxidant damage caused by low Zn to the chloroplast ultrastructure.     

不同磷源对磷高效利用野生大麦根际土壤磷组分的影响

在土培盆栽条件下,以野生大麦磷高效利用基因型IS-22-30、IS-22-25和低效基因型IS-07-07为材料,研究不施磷(CK)、无机磷(KH₂PO₄,Pi)、有机磷(phytate,Po)及二者混合(KH₂PO₄+phytate,Pi+Po)的方式施磷30 mg·kg^-1时,磷高效基因型野生大麦对磷素吸收利用能力及土壤磷组分特征.结果表明： Pi处理野生大麦干物质量和磷素积累量最大,Pi+Po处理其次,Po处理最小,均显著高于CK处理,且磷高效基因型物质生产和磷素吸收能力显著高于磷低效基因型.土壤有效磷在不同磷源处理间差异显著,Pi处理时含量最高,Pi+Po处理次之,且磷高效基因型野生大麦根际有效磷含量显著高于磷低效基因型.磷高效基因型野生大麦根际有效磷呈现亏缺现象,在Pi和Pi+Po处理时亏缺程度较大.根际与非根际土壤无机磷组分含量为Ca₁₀-P>O-P>Fe-P>Al-P>Ca₂-P>Ca₈-P,且其含量随着Pi的增加而增加.各磷源处理下,磷高效基因型野生大麦根际土壤Ca₂-P、Ca₈-P出现亏缺;Pi处理磷高效基因型野生大麦根际土壤Al-P、Fe-P出现富集.土壤中有机磷各组分含量为中活性有机磷>中稳性有机磷、高稳性有机磷>活性有机磷.野生大麦根际土壤活性有机磷和中活性有机磷呈现富集,其富集量在Pi处理时最大;中稳性有机磷和高稳性有机磷呈现亏缺.各磷源处理下,磷高效基因型野生大麦根际土壤活性有机磷含量显著高于磷低效基因型,中稳性有机磷和高稳性有机磷在基因型间差异不显著.Pi缺乏时,磷高效基因型野生大麦活化吸收Ca₂-P、Ca₈-P、Al-P和活性有机磷的能力较强.     

Genetic diversity for mycorrhizal symbiosis and phosphate transporters in rice

Phosphorus(P) is a major plant nutrient and developing crops with higher P-use efficiency is an important breeding goal.In this context we have conducted a comparative study of irrigated and rainfed rice varieties to assess genotypic differences in colonization with arbuscular mycorrhizal(AM) fungi and expression of different P transporter genes.Plants were grown in three different soil samples from a rice farm in the Philippines.The data show that AM symbiosis in all varieties was established after 4 weeks of growth under aerobic conditions and that,in soil derived from a rice paddy,natural AM populations recovered within6 weeks.The analysis of AM marker genes(AM1,AM3,AM14) and P transporter genes for the direct Pi uptake(PT2,PT6) and AM-mediated pathway(PT11,PT13) were largely in agreement with the observed root AM colonization providing a useful tool for diversity studies.Interestingly,delayed AM colonization was observed in the aus-type rice varieties which might be due to their different root structure and might confer an advantage for weed competition in the field.The data further showed that P-starvation induced root growth and expression of the high-affinity P transporter PT6 was highest in the irrigated variety IR66 which also maintained grain yield under P-deficient field conditions.