Different effects of night versus day high temperature on rice quality and accumulation profiling of rice grain proteins during grain filling

High temperature has adverse effects on rice yield and quality. The different influences of night high temperature (NHT) and day high temperature (DHT) on rice quality and seed protein accumulation profiles during grain filling in indica rice ‘9311’ were studied in this research. The treatment temperatures of the control, NHT, and DHT were 28°C/20°C, 27°C/35°C, and 35°C/27°C, respectively, and all the treatments were maintained for 20 days. The result of rice quality analysis indicated that compared with DHT, NHT exerted less effect on head rice rate and chalkiness, whereas greater effect on grain weight. Moreover, the dynamic accumulation change profiles of 61 protein spots, differentially accumulated and successfully identified under NHT and DHT conditions, were performed by proteomic approach. The results also showed that the different suppressed extent of accumulation amount of cyPPDKB might result in different grain chalkiness between NHT and DHT. Most identified isoforms of proteins, such as PPDK and pullulanase, displayed different accumulation change patterns between NHT and DHT. In addition, compared with DHT, NHT resulted in the unique accumulation patterns of stress and defense proteins. Taken together, the mechanisms of seed protein accumulation profiles induced by NHT and DHT during grain filling should be different in rice, and the potential molecular basis is discussed in this study.     

Leaf senescence and grain filling affected by post-anthesis high temperatures in two different wheat cultivars

High temperature is a major factor affecting grain yield and plant senescence in wheat growing regions of central and east China. In this study, two different wheat cultivars, Yangmai 9 with low-grain protein concentration and Xuzhou 26 with high-grain protein concentration, were exposed to different temperature regimes in growth chambers during grain filling. Four day/night temperature regimes of 34°C/22°C, 32°C/24°C, 26°C/14°C, and 24°C/16°C were established to obtain two daily temperatures of 28 and 20°C, and two diurnal day/night temperature differences of 12 and 8°C. Concentration of a lipid peroxidation product malondialdehyde (MDA), activities of the antioxidants superoxide dismutase (SOD) and catalase (CAT), chlorophyll concentration (SPAD) in flag leaves and kernel weight were determined. Results show that activities of SOD and CAT in leaves increased markedly on 14 days after anthesis (DAA) for the high-temperature treatment (34°C/22°C) and then declined. As a result, MDA concentration in leaves increased significantly under high temperature (34°C/22°C and 32°C/24°C). Compared with optimum temperature treatment, high temperature reduced the concentration of soluble protein and SPAD values in flag leaves. Grain-filling rate increased slightly initially, but decreased significantly during late grain filling under high temperature. As a result, final grain weight was reduced markedly under high temperature. Decreases in the activities of SOD and CAT and increases in MDA concentration in leaves were more pronounced with a 12°C of day/night temperature difference when under high temperatures. Kernel weight was higher under 12°C of day/night temperature difference under optimum temperatures (24°C/16°C and 26°C/14°C). The responses to high-temperature regimes appeared to differ between the two wheat cultivars with different grain protein concentrations. It is concluded that a larger diurnal temperature difference hastened the senescence of flag leaves under high-temperature conditions, but retarded senescence under optimum temperature treatments of 26°C/14°C and 24°C/16°C.     

A proteomic analysis of rice seed germination as affected by high temperature and ABA treatment

Seed germination is a critical phase in the plant life cycle, but the specific events associated with seed germination are still not fully understood. In this study, we used two‐dimensional gel electrophoresis followed by mass spectrometry to investigate the changes in the proteome during imbibition of Oryza sativa seeds at optimal temperature with or without abscisic acid (ABA) and high temperature (germination thermoinhibition) to further identify and quantify key proteins required for seed germination. A total of 121 protein spots showed a significant change in abundance (1.5‐fold increase/decrease) during germination under all conditions. Among these proteins, we found seven proteins specifically associated with seed germination including glycosyl hydrolases family 38 protein, granule‐bound starch synthase 1, Os03g0842900 (putative steroleosin‐B), N‐carbamoylputrescine amidase, spermidine synthase 1, tubulin α‐1 chain and glutelin type‐A; and a total of 20 imbibition response proteins involved in energy metabolism, cell growth, cell defense and storage proteins. High temperature inhibited seed germination by decreasing the abundance of proteins involved in methionine metabolism, amino acid biosynthesis, energy metabolism, reserve degradation, protein folding and stress responses. ABA treatment inhibited germination and decreased the abundance of proteins associated with methionine metabolism, energy production and cell division. Our results show that changes in many biological processes including energy metabolism, protein synthesis and cell defense and rescue occurred as a result of all treatments, while enzymes involved in methionine metabolism and weakening of cell wall specifically accumulated when the seeds germinated at the optimal temperature.     

Suppression of phospholipase D genes improves chalky grain production by high temperature during the grain-filling stage in rice

High temperature (HT) during the grain developing stage causes deleterious effects on rice quality resulting in mature grains with a chalky appearance. Phospholipase D (PLD) plays an important role in plants, including responses to environmental stresses. OsPLDα1, α3 and β2-knockdown (KD) plants showed decreased production of chalky grains at HT. HT ripening increased H₂O₂ accumulated in the developing grains. However, the increase was canceled by the knockdown of OsPLDβ2. Expression levels of OsCATA which is one of three rice catalase genes, in developing grains of OsPLDβ2-KD plants at 10 DAF were increased compared with that in vector-controls in HT growth conditions. Overexpression of OsCATA markedly suppressed the production of chalky grains in HT growth conditions. These results suggested that OsPLDβ2 functions as a negative regulator of the induction of OsCATA and is involved in the production of chalky grains in HT growth conditions.     

灌浆期高温和水分逆境对冬小麦籽粒蛋白质和淀粉含量的影响

灌浆期高温和水分逆境是影响小麦籽粒产量和品质的关键气候因子。以扬麦9号、徐州26和豫麦34三个小麦品种为材料,利用人工气候室模拟灌浆期高温和水分胁迫环境,研究了花后高温及温度和水分互作对小麦籽粒蛋白质和淀粉形成的影响。结果表明,高温显著提高了小麦籽粒蛋白质含量及清蛋白、球蛋白和醇溶蛋白含量,但降低了谷蛋白含量,导致麦谷蛋白／醇溶蛋白比值降低。高温显著降低了籽粒总淀粉和支链淀粉含量及支／直比。籽粒蛋白质和淀粉及其组分形成所需的适宜昼夜温差随小麦品质类型而异,但温度水平对籽粒蛋白质和淀粉的影响较温差大。在高温和水分逆境下,温度对籽粒蛋白质和淀粉含量的影响较水分逆境大,且存在显著的互作效应。小麦籽粒蛋白质含量均表现为干旱〉对照〉渍水,以高温干旱最高,适温渍水最低;淀粉含量为对照〉干旱〉渍水,以适温对照最高,而高温渍水最低。高温和水分逆境显著提高了籽粒醇溶蛋白含量而降低了谷蛋白含量及支链淀粉含量,使蛋白质谷／醇比和淀粉支／直比降低,以高温渍水对籽粒蛋白质和淀粉组分的影响最为显著。不同品种之间,高蛋白小麦籽粒蛋白质和组分的形成受高温和水分逆境的影响更大,而低蛋白品种籽粒淀粉形成显著受温度和水分逆境的调节。分析表明,在高温和水分逆境下籽粒蛋白质含量与清蛋白和醇溶蛋白显著正相关,籽粒淀粉含量与谷蛋白、支链淀粉含量及支／直比显著正相关。     

Identification of nutrient partitioning genes participating in rice grain filling by singular value decomposition (SVD) of genome expression data

Background

In order to identify rice genes involved in nutrient partitioning, microarray experiments have been done to quantify genomic scale gene expression. Genes involved in nutrient partitioning, specifically grain filling, will be used to identify other co-regulated genes, and DNA binding proteins. Proper identification of the initial set of bait genes used for further investigation is critical. Hierarchical clustering is useful for grouping genes with similar expression profiles, but decreases in utility as data complexity and systematic noise increases. Also, its rigid classification of genes is not consistent with our belief that some genes exhibit multifaceted, context dependent regulation.

Results

Singular value decomposition (SVD) of microarray data was investigated as a method to complement current techniques for gene expression pattern recognition. SVD's usefulness, in finding likely participants in grain filling, was measured by comparison with results obtained previously via clustering. 84 percent of these known grain-filling genes were re-identified after detailed SVD analysis. An additional set of 28 genes exhibited a stronger grain-filling pattern than those grain-filling genes that were unselected. They also had upstream sequence containing motifs over-represented among grain filling genes.

Conclusions

The pattern-based perspective that SVD provides complements to widely used clustering methods. The singular vectors provide information about patterns that exist in the data. Other aspects of the decomposition indicate the extent to which a gene exhibits a pattern similar to those provided by the singular vectors. Thus, once a set of interesting patterns has been identified, genes can be ranked by their relationship with said patterns.

     

水稻品种灌浆期耐热性的综合评判

建立了一种对水稻品种的灌浆期耐热性进行综合评判的方法.选取籽粒充实度、秕粒率、饱粒密度、垩白米率、垩白度等5个与水稻灌浆期耐热性强弱密切相关的性状,以参评品种这5个性状的耐热系数为指标,应用主成分分析法将5个彼此相关的单项指标转化为2个累计贡献率达到85%以上的相互独立的综合指标,根据每一品种的5个单项性状的耐热系数和综合指标的标准化特征向量求出每一品种2个综合指标的得分.计算每一品种2个综合指标得分的隶属函数值,并以综合指标的贡献率来确定2个综合指标的权重,在此基础上进行加权求和,从而得到每一品种灌浆期耐热性的综合评判值（D值）.采用系统聚类分析方法对各品种的综合评判值（D值）进行数量分类,同时结合生产实际的要求,可把每一品种划归为不同的耐性等级.外部独立样本组的统计检验以及实践验证的结果表明,所建立的综合评判方法具有理想的效果,可以对不同品种灌浆期综合耐热性能的强弱作出客观、准确的评判.     

控制水稻胚乳淀粉合成代谢若干关键酶基因对花后高温的响应表达

以稻米品质温度敏感型的早籼稻品种嘉早935为材料,利用人工气候箱控温试验和实时荧光定量PCR技术,探讨了不同灌浆温度(日均温分别为22和32 ℃)处理下胚乳淀粉分支酶(SBE)、淀粉去分支酶(DBE)和淀粉合酶(SS)的10个同工型基因(sbe1、sbe3、sbe4、pul、isa1、isa2、isa3、Wx、sss1和sss2a)的相对表达量差异及动态变化特征.结果表明: 淀粉合成相关功能基因对水稻灌浆期高温胁迫的响应表达方式存在明显差异,而且因同工型的类型而不同.在高温处理下,sbe1和sbe3的相对表达量显著下降,二者属于SBE类基因中对高温胁迫较敏感的主要同工型;DBE基因中,pul属于高表达的同工型,而且其对高温胁迫响应比isa1、isa2和isa3敏感;在Wx、sss1和sss2a中,sss2a的相对表达量显著低于sss1和Wx, 但sss2a和sss1对高温胁迫响应比Wx敏感,因此二者可能也是高温胁迫对胚乳淀粉结构进行调控的重要位点,尤其在水稻灌浆的中后期发挥重要作用.     

Freezing pattern in apple seeds as affected by the temperature of fruit storage

Storage of apple fruits ( Pyrus malus L. cv. Golden Delicious) at different temperatures (0, 12 and 35°C) markedly altered the pattern of water freezing in the seeds. Higher temperatures of storage brought about a shift from the sequential to the discontinuous type of freezing in seeds, the low temperature exotherm (LTE) being much more pronounced than the high temperature one (HTE). The occurrence of LTE was highly correlated with embryo death. Fruit storage at higher temperatures also caused a decrease in the threshold super cooling temperature of seeds and of naked embryos, removed from the seed coat and endosperm. The decrease was less pronounced in naked embryos than in intact seeds. The results presented show that the frost resistance of apple seeds relies mainly on the supercooling ability of the embryos and is increased by the presence of seed coat and endosperm. The broad peak of the LTE indicates that, in contrast with other seeds and many super cooling organs, massive ice nucleation in apple seeds occurs within the embryo tissues and that extra organ freezing seems to be of less importance. Therefore, the increased super cooling ability of apple seeds, isolated from fruits stored at higher temperatures, seems to rely on those seed properties that protect embryo cells against heterogeneous ice nucleation.     

Free and protein amino acids,and nutrient concentrations in wheat grain as affected by phosphorus nutrition at various salinity levels

Summary The effects of P nutrition under various salinity levels on the protein, amino acids, and nutrients in mature wheat grain were studied. Mexican dwarf wheat (Triticum aestivum L. var Inia) was grown to maturity in solution cultures with variable concentrations of P (0.5-, 5-, 25- and 50 mg P/l) in combination with NaCl at concentrations producing osmotic potentials (_s) of –0.4-, –1.4-, –2.4- and –4.4-bars. All other essential nutrients were present in adequate concentrations for vigorous plant growth.Increasing levels of P in the nutrient solutions tended to decrease the grain yield, N, Cl, protein-glutamic acid,-proline,-leucine,-glycine, and-serine, while P, K, Mg, Zn, Mn and Cu in the grain were increased. The sum of all protein amino acids in the grain decreased as the concentration of P increased in the nutrient solution. The effect of P on the individual and sum of amino acids tended to show peak amounts at the 5.0 mg P/l treatment level.Increased levels of salinity significantly reduced grain yield, N, proteinglutamic acid,-proline,-leucine,-glycine,-serine,-aspartic acid,-alanine, and-phenylalanine in the grain. The sum of the protein amino acids (mol/g dry wt.) was decreased in the grain from plants grown at –4.4 bars salinity level, but not in the grain from plants receiving less saline treatments. The concentrations of free amino acids: serine, aspartic acid, glutamic acid, alanine, and arginine were lower in the grain produced at the –4.4 bars salinity than at –0.4 bars salinity level. The sum of free amino acids (mol/g dry wt.) in the grain were decreased at the highest salinity level as compared with concentrations found for grain produced at lower salinity levels.There were some interactions found between P and salinity on the protein amino acids and nutrients in the grains.The amounts of essential amino acids expressed in mg/g of protein were not significantly affected by the increasing levels of P and salinity in the nutrient solution and they were found in adequate or greater amounts than those recommended by the World Health Organization and FAO.     

The Rc and Rd genes are involved in proanthocyanidin synthesis in rice pericarp

Different colors, such as purple, brown, red and white, occur in the pericarp of rice. Here, two genes affecting proanthocyanidin synthesis in red- and brown-colored rice were elucidated. Genetic segregation analysis suggested that the Rd and A loci are identical, and both encode dihydroflavonol-4-reductase (DFR). The introduction of the DFR gene into an Rcrd mutant resulted in red-colored rice, which was brown in the original mutant, demonstrating that the Rd locus encodes the DFR protein. Accumulation of proanthocyanidins was observed in the transformants by the introduction of the Rd gene into the rice Rcrd line. Protein blot analysis showed that the DFR gene was translated in seeds with alternative translation initiation. A search for the Rc gene, which encodes a transacting regulatory factor, was conducted using available DNA markers and the Rice Genome Automated Annotation System program. Three candidate genes were identified and cloned from a rice RcRd line and subsequently introduced into a rice rcrd line. Brown-colored seeds were obtained from transgenic plants by the introduction of a gene containing the basic helix-loop-helix (bHLH) motif, demonstrating that the Rc gene encodes a bHLH protein. Comparison of the Rc locus among rice accessions showed that a 14-bp deletion occurred only in the rc locus.     

水稻基因组加倍对籽粒大小调控基因表达的影响

水稻是最重要的粮食作物之一,提高水稻产量一直是育种的主要目标。水稻四倍体相对于二倍体具有籽粒变大、粒重增加的特点,研究基因组加倍后籽粒大小基因的调控模式,在育种应用方面具有十分重要的意义。本文以二倍体 -四倍体水稻为材料,分析6个控制籽粒大小基因在幼穗发育中的表达差异,同时结合转基因实验,探讨基因剂量增加对基因表达水平和籽粒大小的影响。结果发现：基因组加倍后,水稻的发育进程不变,但株高增加,叶片变宽,籽粒变大,增大后的籽粒在籼稻表现为长、宽均增加显著,而在粳稻中长度比宽度增加更为明显。进一步分析控制籽粒大小基因的表达差异情况,发现这些基因的表达不仅受发育时期的影响,在籼粳亚种间也明显不同,即受遗传背景的影响。在基因组加倍的情况下,正调控基因GS5、HGW的表达普遍高于对应的二倍体;负调控基因GS3在籼稻D9311中趋于下调或沉默,而在粳稻DBl中趋于上调,GW2在D9311中上调,而在DBl中趋于沉默。通过转基因实验分析负调控基因GW2在二倍体Bl中的表达趋势,发现其在基因剂量线性增加的情况下,表达水平高于二倍体和四倍体,导致其籽粒变小。本研究结果有助于了解水稻中控制籽粒大小的基因在二倍体和四倍体中的表达模式,为高产育种提供理论依据。     

Longevity and temperature response of pollen as affected by elevated growth temperature and carbon dioxide in peanut and grain sorghum

It is important to understand the effects of environmental conditions during plant growth on longevity and temperature response of pollen. Objectives of this study were to determine the influence of growth temperature and/or carbon dioxide (CO₂) concentration on pollen longevity and temperature response of peanut and grain sorghum pollen. Plants were grown at daytime maximum/nighttime minimum temperatures of 32/22, 36/26, 40/30 and 44/34 °C at ambient (350 μmol mol⁻¹) and at elevated (700 μmol mol⁻¹) CO₂ from emergence to maturity. At flowering, pollen longevity was estimated by measuring in vitro pollen germination at different time intervals after anther dehiscence. Temperature response of pollen was measured by germinating pollen on artificial growth medium at temperatures ranging from 12 to 48 °C in incubators at 4 °C intervals. Elevated growth temperature decreased pollen germination percentage in both crop species. Sorghum pollen had shorter longevity than peanut pollen. There was no influence of CO₂ on pollen longevity. Pollen longevity of sorghum at 36/26 °C was about 2 h shorter than at 32/22 °C. There was no effect of growth temperature or CO₂ on cardinal temperatures (T_min, T_opt, and T_max) of pollen in both crop species. The T_min, T_opt, and T_max identified at different growth temperatures and CO₂ levels were similar at 14.9, 30.1, and 45.6 °C, respectively for peanut pollen. The corresponding values for sorghum pollen were 17.2, 29.4, and 41.7 °C. In conclusion, pollen longevity and pollen germination percentage was decreased by growth at elevated temperature, and pollen developed at elevated temperature and/or elevated CO₂ did not have greater temperature tolerance.