Effects of High Temperature and Strong Light Combine Stress on Yield and Quality of Early <i>Indica</i> Rice with Different Amylose Content during Grout Filling

High temperature (HT) accompanied with strong light (SL) often occurs in early indica rice production during grout filling stage in southern China, which accelerates grain ripening. Two indica rice cultivars with different amylose content (Zhongjiazao17, ZJZ17, high amylose content; Xiangzaoxian45, XZX45, low amylose content) were grown under control (CK), HT, and HT+SL conditions during grout filling to determine the effects on grain yield and quality of rice. The results showed that compared with CK, HT and HT+SL significantly reduced the 1000-grain weight and filled grain rate whether in high or low amylose content early indica rice cultivars during grout filling, resulting in a significantly lower grain yield. Meanwhile, HT and HT+SL significantly decreased the milled rice rate, brown rice rate and head rice rate, whereas significantly increased chalky rate and chalky degree; and breakdown decreased and setback, pasting temperature increased in the cultivars, leading to the poor processing, appearance and cooking and eating quality of early indica rice cultivars. Compared with HT, the yield of ZJZ17 was significantly decreased under HT+SL, due to the lower 1000-grain weight. However, the effect of HT+SL on rice quality varied in the cultivars. In general, the yield and rice quality of ZJZ17 were relatively poor under HT+SL. Our results suggested that HT and HT+SL during grout filling had significant damage to the yield and quality of early indica rice cultivars, especially HT+SL, while the high amylose cultivar ZJZ17 showed a higher negative effect under HT+SL.     

The Endosperm-Specific Expression of <i>YUCCA</i> Genes Enhances Rice Grain Filling

Grain filling is a crucial process that affects yield in rice (Oryza sativa L.). Auxin biosynthesis and signaling are closely related to rice yield; therefore, it is important to understand the effects of auxin biosynthesis on rice grain filling to improve crop yield. In this study, we used physiological and molecular strategies to identify the roles of auxin in rice grain filling. Exogenous application of auxin (IAA) or auxin analogues (2, 4-D) to young spikelets and flag leaves improved the seed-setting rate and yield per spike. Furthermore, real-time quantitative PCR assays confirmed that nine members of the OsYUCCA family of auxin biosynthetic genes were upregulated during grain filling, implication that auxin biosynthesis plays a major role in grain development. The specific expression of either Arabidopsis AtYUCCA1 or OsYUCCA2 in the endosperm or leaves resulted in increased expression of OsIAA genes and auxin content of seeds, as well as increased grain filling and seed-setting rate. This result establishes that the auxin content in grains and leaves is important for grain development. Our findings further highlight the potential applications for improving rice yield by elevating targeted gene expression in specific tissues.     

Comparison on physiological adaptation and phosphorus use efficiency of upland rice and lowland rice under alternate wetting and drying irrigation

As one of the most widely promoted water-saving irrigation strategies for rice, alternate wetting and drying irrigation (AWD) can not only save water but also increase mineral nutrient use efficiency. In this study, we compared the growth conditions of four rice varieties (two lowland and two upland varieties) under three irrigation regimes: continuously flooded (CF), alternate wetting and moderate soil drying (AWD15) and alternate wetting and severe soil drying (AWD30). AWD15 and AWD30 enabled the plants to receive fewer irrigation events and less irrigation water than CF, thereby saving both water resources and labor. AWD15 reduced redundant vegetative growth, promoted root growth, and increased the root-shoot ratio and harvest index. AWD15 increased the grain yield, water use efficiency (WUE) and phosphorus use efficiency (PUE) of upland rice and maintained the grain yield while increasing the WUE and PUE of lowland rice. More developed root systems under AWD helped upland rice to maintain a higher water status than lowland rice when plants were subjected to soil drying, which resulted in superior performance in grain yield in upland rice. AWD30 could not reconcile the demands of higher yield and the desire to reduce irrigation water use because it decreased grain yield. The results indicate that AWD15 irrigation of rice can not only increase rice yield and WUE but also enhance PUE, which can potentially reduce the use of phosphorus fertilizers. The results provide theoretical and technical support for improving rice cultivation.     

DDG1 and G Protein α Subunit RGA1 Interaction Regulates Plant Height and Senescence in Rice (<i>Oryza sativa</i>)

Many studies have already shown that dwarfism and moderate delayed leaf senescence positively impact rice yield, but the underlying molecular mechanism of dwarfism and leaf senescence remains largely unknown. Here, using map-based cloning, we identified an allele of DEP2, DDG1, which controls plant height and leaf senescence in rice. The ddg1 mutant displayed dwarfism, short panicles, and delayed leaf senescence. Compared with the wild-type, ddg1 was insensitive to exogenous gibberellins (GA) and brassinolide (BR). DDG1 is expressed in various organs, especially in stems and panicles. Yeast two-hybrid assay, bimolecular fluorescent complementation and luciferase complementation image assay showed that DDG1 interacts with the α-subunit of the heterotrimeric G protein. Disruption of RGA1 resulted in dwarfism, short panicles, and darker-green leaves. Furthermore, we found that ddg1 and the RGA1 mutant was more sensitive to salt treatment, suggesting that DDG1 and RGA1 are involved in regulating salt stress response in rice. Our results show that DDG1/DEP2 regulates plant height and leaf senescence through interacting with RGA1.     

Effects of Ultrasonic Seed Treatment on Rice Performances under the Seawater Irrigation

Irrigation with desalinated seawater is an effective way to use ocean resources and save freshwater resources. However, seawater irrigation would cause yield loss of rice. In order to explore the effects of ultrasonic seed treatment on rice performances under seawater irrigation, the present study was conducted with three irrigation treatments (fresh water (SW0), ten times diluted seawater (SW1%, 0.34% salinity), and five times diluted seawater (SW2%, 0.68% salinity)) and two seed treatments (ultrasonic treated seeds (UT) and untreated seeds (CK)). Compared with SW0 + CK treatment, SW1 + CK and SW2 + CK treatments significantly decreased grain yield by 56.19% and 66.69%, spikelets per panicle by 30.11% and 55.80%, seed-setting rate by 23.05% and 18.87%, and 1000-grain weight by 4.55% and 14.50%, respectively. Seawater irrigation also significantly increased malonaldehyde (MDA) and proline contents and the activities of superoxide dismutase (SOD) and peroxidase (POD). Ultrasonic seed treatment significantly increased the grain number per panicle, seed-setting rate, and grain yield of rice under seawater irrigation. Compared with CK, UT treatment substantially reduced MDA content, SOD activity, and POD activity in SW1 and SW2 conditions. Furthermore, UT treatment significantly increased proline content and down-regulated proline dehydrogenase activity under seawater irrigation. We deduced that ultrasonic seed treatment enhanced the salinity tolerance of rice by inducing the proline accmulation. Our findings indicated that ultrasonic seed treatment could an effective strategy to promote rice productivity under seawater irrigation.     

Proteomic analysis of positive influence of alternate wetting and moderate soil drying on the process of rice grain filling

Rice (Oryza sativa L.) is one of the most important crops in the world to feed ever increasing world population. An increase in output of rice crop per unit is imminent. Alternate wetting and moderate soil drying (AWD) irrigation technology has been recommended as a good practice method to improve grain filling of rice crop at late growing stages. Physiological, molecular and agronomic parameters were adopted to elucidate the role of rice stem and sheath under AWD treatment as compared to the conventional irrigation during the grain filling stage. AWD treatment significantly increased stem and sheath dry weight, carbohydrate reserves and their remobilization to the grain, especially inferior spikelet grains. The results showed that the transport and conversion rate of the stem and sheath photoassimilates increased by 9.87 and 8.37%, respectively. Furthermore, protein expression profiles of the stem and sheath at 10, 20 and 30 days after flowering were analyzed. We examined 220 differentially expressed proteins, and successfully identified 166 proteins, including 71 proteins in the stem and 95 proteins in the sheath involved in thirteen important functional groups. Our results suggest that the AWD treatment at the rice grain filling stage is highly conducive to trigger the mobilization of the N assimilates from leave and root to the stem and sheath, and then promotes to remobilize the reserves to the grain through coordinately expressed proteins involved in photosynthesis, systematic senescence, oxidative stress defense, signal transduction and other metabolisms. This study reveals the metabolic mechanism of the stem and sheath in response to AWD at grain filling stage, and provides theoretical evidence for better quality control and scientific improvement of rice in practice.     

Improvement of rice transformation using bombardment of scutellum-derived calli

Although several reports on rice transformation have been published, producing transgenic plants ofIndica rice varieties is still problematic. We report an improved protocol for transformingIndica rice genotypes. An important agronomic MexicanIndica rice variety, Morelos A-92, was used. Calli derived from scutellum seeds were produced by using auxins and bombarded with 2 vectors, one harboring the reporteruidA gene and the other with thehptII gene conferring hygromycin resistance. The influence of the molar relation of these vectors (uidA-hptIII) in generating callus and plants expressing theuidA reporter gene was analyzed. Selection of bombarded calli was performed under 2 conditions: 50 mg/L of hygromycin with 3 subcultures and 80 mg/L of hygromycin with no subcultures. The best conditions were a 20∶1uidA-hptII molar vector relationship and selection at 80 mg/L of hygromycin, producing 14% of calli expressing GUS. The minimal callus size in regenerating plants was 3 mm. Transformed rice plants were generated with 4.6% efficiency, considering the initial number of bombarded calli. Heredity of theuidA gene behaved as a single locus in transformed rice plants. Homozygous plants were identified in the T₁ generation by means of pollen staining.     

Microalgae Improve the Photosynthetic Performance of Rice Seedlings (<i>Oryza sativa</i> L.) under Physiological Conditions and Cadmium Stress

The aim of this study was to assess the impact of the microalgae Chlorella vulgaris on the rice seedlings at physiological conditions and under cadmium (Cd) stress. We examined the effects of C. vulgaris in the nutrient solution on rice seedlings grown hydroponically in the presence and the absence of 150 μM CdCl₂, using the low (77 K) temperature and pulse amplitude modulated (PAM) chlorophyll fluorescence, P700 photooxidation measurements, photochemical activities of both photosystems, kinetic parameters of oxygen evolution, oxidative stress markers (MDA, H₂O₂ and proline), pigment content, growth parameters and Cd accumulation. Data revealed that the application C. vulgaris not only stimulates growth and improves the functions of photosynthetic apparatus under physiological conditions, but also reduces the toxic effect of Cd on rice seedlings. Furthermore, the presence of the green microalgae in the nutrient solution of the rice seedlings during Cd exposure, significantly improved the growth, photochemical activities of both photosystems, the kinetic parameters of the oxygen-evolving reactions, pigment content and decreased lipid peroxidation, H₂O₂ and proline content. Data showed that the alleviation of Cd-induced effects in rice seedlings is a result of the Cd sorption by microalgae, as well as the reduced Cd accumulation in the roots and its translocation from the roots to the shoots.     

Performance of Soft Rice (<i>Oryza sativa L</i>.) Grown in Early Season in China

Eating quality is of paramount importance to rice (Oryza sativa L.) consumers and soft rice with low amylose content has become popular in China. This study was conducted to evaluate the performance of soft rice grown in the early season (ES) dominated by non-soft rice. Field experiments were conducted in Yongan and Santang, Hunan Province, China from 2016-2018. Results showed that grain amylose content in soft rice cultivars was consistently lower in the ES compared to the late season (LS). The lower grain amylose content in the ES compared to the LS was partly attributed to higher average daily mean temperature during grain filling. No significant relationship was observed between grain yield and seed amylose content in ES rice. Soft rice cultivars produced a similar average grain yield to non-soft rice cultivars in the ES. These results encourage breeders to develop more ES rice cultivars with soft texture to meet the consumer demand for this type of rice.     

Differentiation of Indica-Japonica rice revealed by insertion/deletion (InDel) fragments obtained from the comparative genomic study of DNA sequences between 93-11 ( Indica ) and Nipponbare ( Japonica )

DNA polymorphisms from nucleotide insertion/deletions (InDels) in genomic sequences are the basis for developing InDel molecular markers. To validate the InDel primer pairs on the basis of the comparative genomic study on DNA sequences between an Indica rice 93-11 and a Japonica rice Nipponbare for identifying Indica and Japonica rice varieties and studying wild Oryza species, we studied 49 Indica, 43 Japonica, and 24 wild rice accessions collected from ten Asian countries using 45 InDel primer pairs. Results indicated that of the 45 InDel primer pairs, 41 can accurately identify Indica and Japonica rice varieties with a reliability of over 80%. The scatter plotting data of the principal component analysis (PCA) indicated that: (i) the InDel primer pairs can easily distinguish Indica from Japonica rice varieties, in addition to revealing their genetic differentiation; (ii) the AA-genome wild rice species showed a relatively close genetic relationship with the Indica rice varieties; and (iii) the non-AA genome wild rice species did not show evident differentiation into the Indica and Japonica types. It is concluded from the study that most of the InDel primer pairs obtained from DNA sequences of 93-11 and Nipponbare can be used for identifying Indica and Japonica rice varieties, and for studying genetic relationships of wild rice species, particularly in terms of the Indica-Japonica differentiation. Translated from Journal of Fudan University (Natural Science), 2006, 45(3): 309–315 [译自: 复旦学报(自然科学版)]     

Different isoforms of starch-synthesizing enzymes controlling amylose and amylopectin content in rice (Oryza sativa L.)

Starch, composed of amylose and amylopectin, greatly influences rice cooking and textural quality, which in turn is controlled by various isoforms of several enzymes. Activity of one or more isoforms of starch‐synthesizing enzymes results in various forms of starch structure based on the amylopectin chain length and average external, internal and core chain length distribution and hence results in varying physicochemical and cooking quality. Since the synthesis of starch is highly complex, it is crucial but essential to understand its biosynthetic pathway, starch structure and effects on the physicochemical properties that control eating and cooking quality, and alongside conduct research on gene/QTL mapping for use in marker-assisted selection (MAS) with a view to improve and select cultivars with most desirable range and class of rice starch properties. This article presents the updates on current understanding of the coordination among various enzymes/isoforms towards rice starch synthesis in endosperm and their effect on rice grain physicochemical, cooking and eating qualities. The efforts in identifying regions responsible for these enzymes by mapping the gene/QTLs have provided a glimpse on their association with physicochemical and cooking properties of rice and, hence, improvement is possible by modifying the allelic pattern, resulting in down or nil regulation of a particular enzyme. The clear understanding of the tissue specific coordination between enzyme isoforms and their subsequent effect in controlling eating and cooking properties will enhance the chances to manipulate them for getting desired range of amylose content (AC) and gelatinization temperature (GT) in improved cultivars through combining desired alleles through MAS.     

Map-based cloning of the <Emphasis Type="Italic">ALK</Emphasis> gene,which controls the gelatinization temperature of rice

Gelatinization temperature (GT) is an important parameter for evaluating the cooking and eating quality of rice besides amylose content (AC). The inheritance of the genes affecting GT has been widely studied and is considered to be controlled by a major gene. Here, we report the map-based cloning of rice ALK that encodes the soluble starch synthase II (SSSII). Comparison between the DNA sequences from different rice varieties, together with the results obtained with digestion of the rice seeds in alkali solution, indicates that the base substitutions in coding sequence of ALK may cause the alteration in GT.     

抽穗扬花期不同灌水处理对晚稻抵御低温、产量和生理特性的影响

以超级杂交晚稻品种‘五丰优T025’为材料,设置日排夜灌,灌溉水深4～5 cm(H₁);日排夜灌,灌溉水深8～10 cm(H₂);深水灌溉,保持水深8～10 cm(H₃)等3个处理,以稻田湿润处理水层0～1 cm为对照,研究了抽穗扬花期遭遇低温条件下不同灌水方式和水层深度对双季晚稻生理特性及产量的影响.结果表明: 低温期间,不同灌水处理叶片、土层和冠层温度较对照均有所提高,其中H₂处理增温效果最好.低温胁迫下各处理稻株叶片叶绿素含量、净光合速率、蒸腾速率、叶片气孔导度和胞间CO₂浓度均逐渐降低,其中H₂处理降低幅度最小;H₂处理叶片丙二醛、游离脯氨酸含量上升幅度最小,可溶性蛋白含量高于其余处理,超氧化物歧化酶和过氧化物酶增幅最小,过氧化氢酶下降幅度最小.灌水保温均可达到增产效果,以H₂处理效果最佳,遭遇低温的2014年和2015年第二播期H₂处理分别比对照增产12.9%和13.5%;从产量结构上看,各处理较对照在单株有效穗数、穗长、结实率、千粒重上均有一定的改善,日排夜灌8～10 cm水深处理是增强双季杂交晚稻抽穗扬花期低温抵御能力较为实用的农艺措施.     

Effect of Water Stress Induced by Polyethylene Glycol on Growth,Proline Accumulation in <i>Agave americana</i> L.

The effect of water deficit was determined on both in vitro and soil seedling as well as in cells in suspension of Agave americana L. In order to do the establishment of cells, the formation of callus was induced; for it two auxins were evaluated: 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-mino-3,5,6-trichloropicolinic acid (picloram) at three concentrations (0.25, 0.5 and 0.75 mg L⁻¹) in three explants (leaf, root and meristems) cultured in MS semisolid medium. The callogenesis response was related to the type and section of the explant, as well as the regulator used, and a cell suspension was established using 0.5 mg L⁻¹ naphthaleneacetic acid (NAA) + 0.5 mg L⁻¹ Benzylaminopurine (BAP). Seedlings were exposed to polyethyleneglycol (15% and 30% w/v) with a water potential of −0.87 and −2.67 MPa, respectively, under soil conditions. Water stress was applied through restricted irrigation. Fresh weight, root system growth, and chlorophyll concentration were some of the parameters that were affected by the effect of water deficit on A. americana L. Chlorophyll concentration values were significantly decreased by 15 at 30% PEG (19.6 SPAD units) compared to the control treatment. In in vitro plants, the highest concentration of proline was found in the roots, being the treatment with 30% polyethylene glycol where the highest concentration of this osmoregulator was obtained (62.5 mg g⁻¹ DW). Under restricted irrigation conditions, an increase in proline concentration was observed both in the aerial part (2.2 µg 100 g⁻¹ DW) and in the root system (1.8 µg 100 g⁻¹ DW). However, the concentrations found were approximately ten times greater, less than those found under in vitro conditions. Therefore, the accumulation of proline can be considered an indicator of stress in Agave Americana L. growth in vitro.     

Identification of a Novel <i>OsCYP2</i> Allele that Was Involved in Rice Response to Low Temperature Stress

Cyclophilin (CYP) plays an important role in plant response to stress, and OsCYP2, one gene of cyclophlilin family, is involved in auxin signal transduction and stress signaling in rice. However, the mechanism that OsCYP2 is involved in rice response to low temperature is still unclear. We identified a new OsCYP2 allelic mutant, lrl3, with fewer lateral roots, and the differences in shoot height, primary root length and adventitious root length increased with the growth process compared to the wild-type plant. Auxin signaling pathway was also affected and became insensitive to gravity. The transgenic rice plants with over-expression of OsCYP2 were more tolerant to low temperature than the wild-type plants, suggesting that OsCYP2 was involved in the low temperature response in rice. In addition, OsCYP2 negatively regulated the expression of OsTPS38, a terpene synthase gene, and was dependent on the OsCDPK7-mediated pathway in response to low temperature stress. OsTPS38- overexpressed transgenic line ox-2 was more sensitive to low temperature. Therefore, OsCYP2 may negatively regulate OsTPS38 through an OsCDPK7-dependent pathway to mediate the response to low temperature in rice. These results provide a new basis for auxin signaling genes to regulate rice response to low temperature stress.     

Differential Responses of <i>NHX1</i> and <i>SOS1</i> Gene Expressions to Salinity in two <i>Miscanthus sinensis</i> Anderss. Accessions with Different Salt Tolerance

The lignocellulosic crop Miscanthus spp. has been identified as a good candidate for biomass production. The responses of Miscanthus sinensis Anderss. to salinity were studied to satisfy the needs for high yields in marginal areas and to avoid competition with food production. The results indicated that the relative advantages of the tolerant accession over the sensitive one under saline conditions were associated with restricted Na⁺ accumulation in shoots. Seedlings of two accessions (salt-tolerant ‘JM0119’ and salt-sensitive ‘JM0099’) were subjected to 0 (control), 100, 200, and 300 mM NaCl stress to better understand the salt-induced biochemical responses of genes involved in Na⁺ accumulation in M. sinensis. The adaptation responses of genes encoding for Na⁺ /H⁺ antiporters, NHX1 and SOS1 to NaCl stress were examined in JM0119 and JM0099.The cDNA sequences of genes examined were highly conserved among the relatives of M. sinensis based on the sequencing on approximate 600 bp-long cDNA fragments obtained from degenerate PCR. These salt-induced variations of gene expression investigated by quantitative real-time PCR provided evidences for insights of the molecular mechanisms of salt tolerance in M. sinensis. The expression of NHX1 was up-regulated by salt stress in JM0119 shoot and root tissues. However, it was hardly affected in JM0099 shoot tissue except for a significant increase at the 100 mM salt treatment, and it was salt-suppressed in the JM0099 root tissue. In the root tissue, the expression of SOS1 was induced by the high salt treatment in JM0119 but repressed by all salt treatments in JM0099. Thus, the remarkably higher expression of NHX1 and SOS1 were associated with the resistance to Na⁺ toxicity by regulation of the Na⁺ influx, efflux, and sequestration under different salt conditions.     

冬季覆盖作物秸秆还田对双季稻田根际土壤微生物群落功能多样性的影响

微生物群落功能多样性是土壤质量变化重要的指标,不同作物类型的秸秆还田措施对土壤微生物群落功能多样性具有明显的影响。以位于双季稻主产区不同冬季覆盖作物-双季稻种植模式大田定位试验田为研究对象,以冬闲-双季稻种植模式为对照(CK),应用Biolog-GN技术开展黑麦草-双季稻(Ry)、紫云英-双季稻(Mv)、油菜-双季稻(Ra)和马铃薯-双季稻(Po)种植模式条件下不同冬季覆盖作物秸秆还田后对双季稻田根际土壤微生物功能多样性影响的研究。研究结果表明,早稻和晚稻成熟期,与CK处理相比,冬季覆盖作物秸秆还田处理增加了稻田土壤碳源平均颜色变化率(AWCD),以Po处理AWCD均为最高,均显著高于Ry和CK处理。不同冬季覆盖作物秸秆还田处理土壤微生物代谢多样性指数表现出明显的差异,早稻成熟期,Po处理的Richness、Shannon和McIntosh指数均为最高,其次为Ry、Mv和Ra处理,CK处理最低;晚稻成熟期,各处理的Richness、Shannon和McIntosh指数大小顺序均表现为PoRaMvRyCK。土壤微生物碳源利用的主成分分析结果表明,各冬季覆盖作物秸秆还田处理根际土壤微生物利用的主要碳源为氨基酸类和糖类物质,不同处理间碳源利用类型有差异。冬季覆盖作物秸秆还田措施有利于提高双季稻田根际土壤微生物对碳源的利用能力、物种丰富度和均匀度。