Relationship between leaf photosynthetic function at grain filling stage and yield in super high-yielding hybrid rice (Oryza sativa. L)

The characteristics of dry matter production before and after heading and the relationships between photosynthesis of flag leaves and dry matter accumulation in panicles were investigated on super high-yielding rice cv. Xieyou 9308 (the yield of up to 12 t/ha) with rice cv. Xieyou 63 as a control. The results showed that (i) the capacity of dry matter production before and after heading in Xieyou 9308, i.e. biomass and daily dry matter production, was remarkably higher than that in Xieyou 63, especially after heading; (ii) CO2 assimilation capacity in flag leaves in Xieyou 9308, namely Leaf Source Capacity (LSC), was also significantly higher than that in Xieyou 63, and the supply of photosynthate in leaves and the demand of grain filling were completely synchronous in Xieyou 9308, but photosynthetic function in flag leaves in Xieyou 63 declined sharply 20 days after heading and it was not enough to meet the demand of grain filling. These results confirmed that high efficient photosynthetic function in     

Relationship between leaf photosynthetic function at grain filling stage and yield in super high-yielding hybrid rice ( Oryza sativa . L)

The characteristics of dry matter production before and after heading and the relationships between photosynthesis of flag leaves and dry matter accumulation in panicles were investigated on super high-yielding rice cv. Xieyou 9308 (the yield of up to 12 t/ha) with rice cv. Xieyou 63 as a control. The results showed that (i) the capacity of dry matter production before and after heading in Xieyou 9308, i.e. biomass and daily dry matter production, was remarkably higher than that in Xieyou 63, especially after heading; (ii) CO₂ assimilation capacity in flag leaves in Xieyou 9308, namely Leaf Source Capacity (LSC), was also significantly higher than that in Xieyou 63, and the supply of photosynthate in leaves and the demand of grain filling were completely synchronous in Xieyou 9308, but photosynthetic function in flag leaves in Xieyou 63 declined sharply 20 days after heading and it was not enough to meet the demand of grain filling. These results confirmed that high efficient photosynthetic function in leaves after heading and its complete synchronization with grain filling are the key approaches to super high yield of rice.     

Cadmium-induced changes in antioxidant enzyme activities in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L. cv. Dongjin)

We studied how the relationship between cadmium (Cd) toxicity and oxidative stress influenced the growth, photosynthetic efficiency, lipid peroxidation, and activity of ntioxidative enzymes in the roots and leaves of rice(Oryza sativa L Dongjin). Plants were exposed to Cd for 21 d. Both seedling growth and photosynthetic efficiency decreased gradually with increasing cadmium concentrations. Lipid peroxidation increased slowly in both roots and leaves, causing oxidative stress. However, each tissue type responded differently to Cd concentrations with regard to the induction/ inhibition of antioxidative enzymes. The activity of Superoxide dismutase (SOD) increased in both roots and leaves. Ascorbate peroxidase (APX) activity increased in leaves treated with up to 0.25 μM Cd, then decreased gradually at higher concentrations. In contrast, APX activity in roots increased and remained constant between 0.25 and 25 μM Cd. Enhanced peroxidase (POD) activity was recorded for treatments with up to 25/M Cd, gradually decreasing at higher concentrations in the leaves but remaining unchanged in the roots. Catalase (CAT) activity increased in the roots, but decreased in the leaves, whereas the activity of glutathione reductase (GR) was enhanced in both roots and leaves, where it remained elevated at higher Cd concentrations. These results suggest that rice seedlings tend to cope with free radicals generated by Cd through coordinated, enhanced activities of the antioxidative enzymes involved in detoxification.     

Loss of function of <Emphasis Type="Italic">OsMADS34</Emphasis> leads to large sterile lemma and low grain yield in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Rice (Oryza sativa L.) is one of the most important food crops, especially in Asia. The spikelet is a characteristic structure of grass inflorescences that determines crop output. However, the molecular mechanism that controls spikelet development and grain yield in rice remains unclear. In this study, we isolated a new osmads34 allelic mutant (i.e., osmads34-t). The osmads34-t mutant showed more primary branch numbers, short panicles, and long sterile lemmas. The sterile lemmas were transformed into the lemmas and had the lemma identity in the osmads34-t mutant, suggesting that the sterile lemma and lemma are homologous organs. Additionally, osmads34-t displayed smaller grains on its secondary branches of panicles and a lower seed-setting rate. These results suggest that OsMADS34 plays an important role in determination of grain size and yield in rice. OsMADS34 was expressed in tested organs and tissues, and its green fluorescent protein (GFP) signal was located in the nucleus. The result of this study will be used to understand the identity of unique organs in grass spikelets and may improve grain yield in breeding practice.     

Simple sequence repeat analyses of interspecific hybrids and MAALs of <Emphasis Type="Italic">Oryza </Emphasis><Emphasis Type="Italic">officinalis</Emphasis> and <Emphasis Type="Italic">Oryza sativa</Emphasis>

Wild rice is a valuable resource for the genetic improvement of cultivated rice (Oryza sativa L., AA genome). Molecular markers are important tools for monitoring gene introgression from wild rice into cultivated rice. In this study, Simple sequence repeat (SSR) markers were used to analyze interspecific hybrids of O. sativa-O. officinalis (CC genome), the backcrossing progenies and the parent plants. Results showed that most of the SSR primers (335 out of 396, 84.6%) developed in cultivated rice successfully amplified products from DNA samples of wild rice O. officinalis. The polymorphism ratio of SSR bands between O. sativa and O. officinalis was as high as 93.9%, indicating differences between the two species with respect to SSRs. When the SSR markers were applied in the interspecific hybrids, only a portion of SSR primers amplified O. officinalis-specific bands in the F(1) hybrid (52.5%), BC(1) (52.5%), and MAALs (37.0%); a number of the bands disappeared. Of the 124 SSR loci that detected officinalis-specific bands in MAAL plants, 96 (77.4%) showed synteny between the A and C-genomes, and 20 (16.1%) showed duplication in the C-genome. Sequencing analysis revealed that indels, substitution and duplication contribute to the diversity of SSR loci between the genomes of O. sativa and O. officinalis.     

The parameters of grain filling and yield components in common wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) and durum wheat (<Emphasis Type="Italic">Triticum turgidum</Emphasis> L. var. <Emphasis Type="Italic">durum</Emphasis>)

Final grain dry weight, a component of yield in wheat, is dependent on the duration and the rate of grain filling. The purpose of the study was to compare the grain filling patterns between common wheat, (Triticum aestivum L.), and durum wheat, (Triticum turgidum L. var. durum), and investigate relationships among grain filling parameters, yield components and the yield itself. The most important variables in differentiating among grain filling curves were final grain dry weight (W) for common wheat genotypes and grain filling rate (R) for durum wheat genotypes; however, in all cases the sets of variables important in differentiating among grain filling curves were extended to either two or all three parameters. Furthermore, in one out of three environmental conditions and for both groups of genotypes, the most important parameter in the set was grain filling duration (T). It indicates significant impact of environmental conditions on dry matter accumulation and the mutual effect of grain filling duration and its rate on the final grain dry weight. The medium early anthesis date could be associated with further grain weight and yield improvements in wheat. Grain filling of earlier genotypes occurs in more temperate environments, which provides enough time for gradual grain fill and avoids the extremes of temperature and the stress of dry conditions.     

Iron homeostasis in tropical <Emphasis Type="Italic">indica</Emphasis> rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) cultivars having contrasting grain iron concentration

Iron homeostasis was studied in two tropical indica rice cultivars viz. Sharbati (high Fe) and Lalat (low Fe) having contrasting grain Fe concentration. Plants were hydroponically grown with 5 concentrations of Fe (0.05, 2, 5, 15, 50 mg L^?1) till maturity. The effect of incremental Fe treatment on the plant was followed by analyzing accumulation of ferritin protein, activities of aconitase enzyme, enzymes of anti-oxidative defense and accumulation of hydrogen peroxide and ascorbic acid. Plant growth was adversely affected beyond 15 mg L^?1 of Fe supplementation and effects of Fe stress (both deficiency and excess) were more apparent on the high Fe containing cultivar Sharbati than the low Fe containing Lalat. Level of ferritin protein and aconitase activity increased up to 5 mg L^?1 of Fe concentration. Lalat continued to synthesize ferritin protein at much higher Fe level than Sharbati and the cultivar also had higher activities of peroxidase, superoxide dismutase and glutathione reductase. It was concluded that the tolerance of Lalat to Fe stress was because of its higher intrinsic ability to scavenge free radicals of oxidative stress for possessing higher activity of antioxidative enzymes. This, together with its capacity to sequester the excess Fe in ferritin protein over a wider range of Fe concentrations made it more tolerant to Fe stress.     

Characterisation of a novel quantitative trait locus, <Emphasis Type="Italic">GN4</Emphasis>-<Emphasis Type="Italic">1</Emphasis>, for grain number and yield in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Key message

A novel QTL for grain number, GN4-1, was identified and fine-mapped to an ~ 190-kb region on the long arm of rice chromosome 4.

Abstract

Rice grain yield is primarily determined by three components: number of panicles per plant, grain number per panicle and grain weight. Among these traits, grain number per panicle is the major contributor to grain yield formation and is a crucial trait for yield improvement. In this study, we identified a major quantitative trait locus (QTL) responsible for rice grain number on chromosome 4, designated GN4-1 (a QTL for Grain Number on chromosome 4), using advanced segregating populations derived from the crosses between an elite indica cultivar ‘Zhonghui 8006’ (ZH8006) and a japonica rice ‘Wuyunjing 8’ (WYJ8). GN4-1 was delimited to an ~ 190-kb region on chromosome 4. The genetic effect of GN4-1 was estimated using a pair of near-isogenic lines. The GN4-1 gene from WYJ8 promoted accumulation of cytokinins in the inflorescence and increased grain number per panicle by ~ 17%. More importantly, introduction of the WYJ8 GN4-1 gene into ZH8006 increased grain yield by ~ 14.3 and ~ 11.5% in the experimental plots in 2014 and 2015, respectively. In addition, GN4-1 promoted thickening of the culm and may enhance resistance to lodging. These results demonstrate that GN4-1 is a potentially valuable gene for improvement of yield and lodging resistance in rice breeding.

     

Transposition behavior of nonautonomous a <Emphasis Type="Italic">hAT</Emphasis> superfamily transposon <Emphasis Type="Italic">nDart</Emphasis> in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Transposable elements (TEs) have a significant impact on the evolution of gene function and genome structures. An endogenous nonautonomous transposable element nDart was discovered in an albino mutant that had an insertion in the Mg-protoporphyrin IX methyltransferase gene in rice. In this study, we elucidated the transposition behavior of nDart, the frequency of nDart transposition and characterized the footprint of nDart. Novel independent nDart insertions in backcrossed progenies were detected by DNA blotting analysis. In addition, germinal excision of nDart occurred at very low frequency compared with that of somatic excision, 0–13.3%, in the nDart1-4(3-2) and nDart1-A loci by a locus-specific PCR strategy. A total of 253 clones from somatic excision at five nDart loci in 10 varieties were determined. nDart rarely caused deletions beyond target site duplication (TSD). The footprint of nDart contained few transversions of nucleotides flanking to both sides of the TSD. The predominant footprint of nDart was an 8-bp addition. Precise excision of nDart was detected at a rate of only 2.2%, which occurred at two loci among the five loci examined. Furthermore, the results in this study revealed that a highly conserved mechanism of transposition is involved between maize Ac/Ds and rice Dart/nDart, which are two-component transposon systems of the hAT superfamily transposons in plant species.     

Prediction of hybrid grain yield performances in Indica Rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) with effect-increasing loci

One hundred and fifty-one rice hybrids produced in two sets of half-dialell crosses and their parents (13 cytoplasmic male sterile lines and 19 restorers) were used to predict the F₁ performances of seven yield traits through the parental genetic distances (GD) based on SSR markers. The positive loci (PL) and effect-increasing loci (IL), which were screened from SSR polymorphic loci by the F₁ traits of 32 parents, together with total loci (TL), were utilized to estimate parental GD and the models were found to predict the traits of hybrids derived from different parents, fixed parents, and different environments, respectively. The results were as follows: (1) 550 polymorphic loci were detected from 174 SSR markers: a dendrogram based on these loci could separate all the sterile and restorer lines used in the present study, which indicated that parental genetic diversity of F₁ was large; (2) the correlations between F₁ traits and parental GDs based on IL ranged from 0.61 to 0.87 with a mean of 0.76, and they were higher than those on TL or on PL; (3) predictions based on IL for F₁ traits (except grain weight per plant) derived from different environments were ideal, but worse for F₁ traits derived from different parents; and (4) IL was more effective than TL and PL in predicting traits of F₁ with fixed parents, and predictions for fixed restorer combinations were more effective than those for fixed sterile line combinations. These results should facilitate molecular prediction for hybrid yield and other traits by means of both elite sterile and restorer lines.     

Molecular characterization the <Emphasis Type="Italic">YABBY</Emphasis> gene family in <Emphasis Type="Italic">Oryza sativa</Emphasis> and expression analysis of <Emphasis Type="Italic">OsYABBY1</Emphasis>

Members of the YABBY gene family have a general role that promotes abaxial cell fate in a model eudicot, Arabidopsis thaliana. To understand the function of YABBY genes in monocots, we have isolated all YABBY genes in Oryza sativa (rice), and revealed the spatial and temporal expression pattern of one of these genes, OsYABBY1. In rice, eight YABBY genes constitute a small gene family and are classified into four groups according to sequence similarity, exon-intron structure, and organ-specific expression patterns. OsYABBY1 shows unique spatial expression patterns that have not previously been reported for other YABBY genes, so far. OsYABBY1 is expressed in putative precursor cells of both the mestome sheath in the large vascular bundle and the abaxial sclerenchyma in the leaves. In the flower, OsYABBY1 is specifically expressed in the palea and lemma from their inception, and is confined to several cell layers of these organs in the later developmental stages. The OsYABBY1-expressing domains are closely associated with cells that subsequently differentiate into sclerenchymatous cells. These findings suggest that the function of OsYABBY1 is involved in regulating the differentiation of a few specific cell types and is unrelated to polar regulation of lateral organ development.     

Response of antioxidant enzymes in rice (<Emphasis Type="Italic">Oryza sauva</Emphasis> L. cv. Dongjin) under mercury stress

We studied the effects of different concentrations of mercury (0.0 to 100 μM) on growth and photosynthetic efficiency in rice plants treated for 21 d. In addition, we investigated how this metal affected the malondialdehyde (MDA) content as well as the activity of five antioxidant enzymes — superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), guaiacol peroxidase (POD), and catalase (CAT). Photosynthetic efficiency (Fμ/F_m) and seedling growth decreased as the concentration of Hg was increased in the growth media. Plants also responded to Hg-induced oxidative stress by changing the levels of their antioxidative enzymes. Enhanced lipid peroxidation was observed in both leaves and roots that had been exposed to oxidative stress, with leaves showing higher enzymatic activity. Both SOD and APX activities increased in treatments with up to 50 μM Hg, then decreased at higher concentrations. In the leaves, both CAT and POD activities increased gradually, with CAT levels decreasing at higher concentrations. In the roots, however, CAT activity remained unchanged while that of POD increased a bit more than did the control for concentrations of up to 10 μM Hg. At higher Hg levels, both CAT and POD activities decreased. GR activity increased in leaves exposed to no more than 0.25 μM Hg, then decreased gradually. In contrast, its activity was greatly inhibited in the roots. Based on these results, we suggest that when rice plants are exposed to different concentrations of mercury, their antioxidative enzymes become involved in defense mechanisms against the free radicals that are induced by this stress.     

Functional markers for <Emphasis Type="Italic">xa5</Emphasis>-mediated resistance in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis>, L.)

The recent cloning of several agronomically important genes has facilitated the development of functional markers. These markers reside within the target genes themselves and can be used with great reliability and efficiency to identify favorable alleles in a breeding program. Bacterial blight (BB) is a severe rice disease throughout the world that is controlled primarily through use of resistant cultivars. xa5 is a race-specific, recessive gene mediating resistance to BB. It is widely used in rice breeding programs throughout the tropics. Due to its recessive nature, phenotypic selection for xa5-mediated resistance is both slow and costly. Previously, marker assisted selection (MAS) for this resistance gene was not efficient because it involved markers that were only indirectly linked to xa5 and ran the risk of being separated from the trait by recombination. Recently, the cloning of the gene underlying this trait made it possible to develop functional markers. Here we present a set of CAPS markers for easy, quick and direct identification of cultivars or progeny carrying xa5-mediated resistance and provide evidence that these markers are 100% predictive of the presence of the xa5 allele. These markers are expected to enhance the reliability and cost-effectiveness of MAS for xa5-mediated resistance.     

Knockout of a starch synthase gene <Emphasis Type="Italic">OsSSIIIa</Emphasis>/<Emphasis Type="Italic">Flo5</Emphasis> causes white-core floury endosperm in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

To elucidate the role of SSIIIa during starch synthesis in rice (Oryza sativa L.) endosperm, we characterized null mutants of this gene, generated by T-DNA insertions. Scanning electron microscope (SEM) analysis revealed that the starch granules in these mutants are smaller and rounder compared with the wild type controls, and that the mutant endosperm is characterized by a loosely packed central portion exhibiting a floury-like phenotype. Hence, the OsSSIIIa (Oryza sativa SSIIIa) mutations are referred to as white-core floury endosperm 5-1 (flo5-1) and flo5-2. Based upon their X-ray diffraction patterns, the crystallinity of the starch in the flo5 mutant endosperm is decreased compared with wild type. Through determination of the chain-length distribution of the mutant endosperm starch, we found that flo5-1 and flo5-2 mutants have reduced the content of long chains with degree of polymerization (DP) 30 or greater compared with the controls. This suggests that OsSSIIIa/Flo5 plays an important role in generating relatively long chains in rice endosperm. In addition, DP 6 to 8 and DP 16 to 20 appeared to be reduced in endosperm starch of flo5-1 and flo5-2, whereas DP 9 to 15 and DP 22 to 29 were increased in these mutants. By the use of differential scanning calorimetry (DSC), the gelatinization temperatures of endosperm starch were found to be 1–5°C lower than those of the control. We propose a distinct role for OsSSIIIa/Flo5 and the coordinated action of other SS isoforms during starch synthesis in the seed endosperm of rice.     

Fine mapping of <Emphasis Type="Italic">S31</Emphasis>, a gene responsible for hybrid embryo-sac abortion in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Partial abortion of female gametes and the resulting semi-sterility of indica × japonica inter-subspecific rice hybrids have been ascribed to an allelic interaction, which can be avoided by the use of wide compatibility varieties. To further understand the genetic mechanism of hybrid sterility, we have constructed two sets of hybrids, using as male parent either the typical japonica variety Asominori, or the wide compatibility variety 02428; and as female, a set of 66 chromosome segment substitution lines in which various chromosomal segments from the indica variety IR24 have been introduced into a common genetic background of Asominori. Spikelet semi-sterility was observed in hybrid between CSSL34 and Asominori, which is known to carry the sterility gene S31 (Zhao et al. in Euphytica 151:331–337, 2006). Cytological analysis revealed that the semi-sterility of the CSSL34 × Asominori hybrid was caused primarily by partial abortion of the embryo sac at the stage of the mitosis of the functional megaspore. A population of 1,630 progeny of the three-way cross (CSSL34 × 02428) × Asominori was developed to map S31. Based on the physical location of linked molecular markers, S31 was thereby delimited to a 54-kb region on rice chromsome 5. This fragment contains eight predicted open reading frames, four of which encode known proteins and four putative proteins. These results are relevant to the map-based cloning of S31, and the development of marker-assisted transfer of non-sterility allele inducing alleles to breeding germplasm, to allow for a more efficient exploitation of heterosis in hybrid rice.     

Genetic transformation of NERICA,interspecific hybrid rice between <Emphasis Type="Italic">Oryza glaberrima</Emphasis> and <Emphasis Type="Italic">O. sativa</Emphasis>, mediated by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>

We developed an efficient gene transfer method mediated by Agrobacterium tumefaciens for introgression of new rice for Africa (NERICA) cultivars, which are derivatives of interspecific hybrids between Oryza glaberrima Steud. and O. sativa L. Freshly isolated immature embryos were inoculated with A. tumefaciens LBA4404 that harbored binary vector pBIG-ubi::GUS or pIG121Hm, which each carried a hygromycin-resistance gene and a GUS gene. Growth medium supplemented with 500 mg/l cefotaxime and 20 mg/l hygromycin was suitable for elimination of bacteria and selection of transformed cells. Shoots regenerated from the selected cells on MS medium containing 20 g/l sucrose, 30 g/l sorbitol, 2 g/l casamino acids, 0.25 mg/l naphthaleneacetic acid, 2.5 mg/l kinetin, 250 mg/l cefotaxime, and 20 mg/l hygromycin. The shoots developed roots on hormone-free MS medium containing 30 mg/l hygromycin. Integration and expression of the transgenes were confirmed by PCR, Southern blot analysis, and histochemical GUS assay. Stable integration, expression, inheritance, and segregation of the transgenes were demonstrated by molecular and genetic analyses in the T₀ and T₁ generations. Most plants were normal in morphology and fertile. The transformation protocol produced stable transformants from 16 NERICA cultivars. We also obtained transformed plants by inoculation of calluses derived from mature seeds, but the frequency of transformation was lower and sterility was more frequent.