Molecular Cloning and Characterization of Citrate Synthase Gene in Rice （ Oryza sativa）

The full-length OsCS encoding citrate synthase was isolated from rice （Oryza sativa L. subsp, japonica）, OsCS is 1477-bp long and encodes a 474 amino acid polypeptide, Its putative protein sequence is highly identical to Daucus carota, Nicotiana tabacum Beta vulgaris subsp., Arabidopsis thaliana, and Citrus junos （〉70%）. The deduced amino-terminal sequence of OsCS showes characteristics of mitochondrial targeting signal. Southern blot analysis using ORF of the OsCS as the probe indicated that this gene exists in multiple copies in rice genome. The band with predicated size of 82 kD was detected by Western blot after being induced by 0,4 mmol/L IPTG.     

Aboveground Characteristics, Yield Potential and Drought Tolerance in “Konyu” Potato Cultivars with Large Root Mass

To improve drought tolerance of potato, we bred four cultivars with large root mass and registered them as Konyu-1 to Konyu-4 in 2007. In this paper, we present the yield performance and the related aboveground characteristics of three Konyu cultivars and cv. Konafubuki (small root mass, one of the parents of these Konyu cultivars) in irrigated and droughted fields for 3 years. Consistent with our previous reports, we observed that the yield reduction in droughted fields relative to irrigated fields was less in Konyu cultivars than in Konafubuki. Konyu-2 showed the highest yield in both fields in all three experimental years. Konyu-2 and Konafubuki had a relatively high harvest index (HI) as a result of lower dry weight partitioning to the shoot regardless of water treatment. This characteristic contributed to the higher yield in the irrigated field in both cultivars compared with the other cultivars. In addition, Konyu-2 had a higher leaf/stem ratio coupled with a lower number of branches than the other cultivars. In the year of the most severe drought, the leaf area index remained below 3 (optimum value for intercept radiation) in Konafubuki, but not in Konyu-2 because of its high leaf/stem ratio. The present results suggest that in addition to large root mass, high HI coupled with high leaf/stem ratio with low number of branches may contribute to achieve high and stable yields in drought prone environments.     

Mapping a Novel Gene of Cold Tolerance at Booting Stage by Using Near-Isogenic Lines in japonica Rice

Genetic analysis showed that cold tolerance at booting stage of near-isogenic lines （NILs） of Kunmingxiaobaigu was controlled by a gene with large phenotypic variance. One hundred and sixty-four simple sequence repeats （SSR） distributed over 12 chromosomes were used to screen polymorphism between Towata （recurrent parent, RP） and near-isogenic line pool （NILP）, and two SSR markers at the long arm of chromosome 5 showed polymorphism in comparison with RP genome. Of the two markers, RM31 was found possibly linked with the cold tolerance gene at booting stage through one-way ANOVA. Twelve SSR markers around RM31 were then used to detect polymorphism between RP and NIL, and only RM7452 had polymorphism. The gene of cold tolerance at booting stage was further mapped on chromosome 5 between RM7452 and RM31 with genetic distances of 4.8 cM and 8.0 cM, respectively. This gene explained 10.50% of phenotypic variance and 5.10% of phenotypic variance of fully filled grains, and was tentatively designated as Ctb（t）.     

A Study on Tolerance to Pretilachlor in Eight Species of Echinochloa

Height of seedlings, length of leaf and coleoptile cells, and α-amylase activities of eight species or varieties of barnyardgrass (Echinochloa) were measured after seeds being treated with pretilachlor. There existed obvious differences in the tested traits among the materials. The inhibition rate of pretilachlor to Echinochloa crusgalli (L.) Beavu. var. mitis (Pursh.) Peterm was the lowest, and that to E. colonum (L.) Link was the highest among the eight species or varieties of barnyardgrass. It suggested that the different species or varieties of barnyardgrass possess variable tolerance to pretilachlor.     

Characterization of a Maize Sucrose–phosphate Synthase Protein and Its Effect on Carbon Partitioning in Transgenic Rice Plants

Abstract

We obtained transgenic rice (Oryza sativa L. cv. Nipponbare) plants with the gene for maize sucrose-phosphate synthase (EC 2.4.1.14, SPS). Some of the transgenic plants over-expressed maize SPS (over-expressing plants) and some had reduced levels of native SPS protein (co-suppressed plants). There was a positive correlation between the amounts of maize SPS protein and SPS activities. However, apparent Km values for uridine diphosphoglucose (UDPG) were higher in over-expressing plants than in control rice plants. These results suggest that overproduced maize SPS protein was not fully activated. The sucrose contents did not differ significantly between control and over-expressing rice plants, but they were lower in co-suppressed plants than in control plants. The starch contents were negatively and the sucrose/starch ratios were positively correlated with SPS activities. Thus, carbon partitioning in the transgenic rice was changed, even though rice is predominantly a sucrose-former.     

A One-Step PCR Method for Detecting the First Base of Splice Donor of Wx Intron 1 in Rice

A new method of one-step PCR was devised for detecting the first nucleotide in the splice donor site of Wx intron 1. compared to the regular PCR-AccⅠmethod, the method can produce the same result for detecting +1 nucleotide of Wx intron 1. The reliability of the new method was confirmed with 30 rice varieties. The new technique is more convenient and cheaper than the regular PCR-AccⅠmethod, and could be widely deploded in rice molecular marker assistant selection.     

Super Rice Cropping Will Enhance Rice Yield and Reduce CH4 Emission： A Case Study in Nanjing,China

A pot experiment was performed to learn the differences in plant productivity and OH4 emission between two rice cultivars, super rice variety Ningjing 1 and traditional variety Zhendao 11, which were currently commercially appUed in Nanjing, China. Similar seasonal changes of CH4 emission fluxes and soil solution CH4 contents were found between the tested cultivars. Although there was no significant difference in plant biomass production between the cultivars, the grain yield of Ningjing 1 was significantly higher by 35.0% （P 〈 0.05） than that of Zhendao 11, whereas the total CH4 emission from Ningjing 1 was 35.2% lower （P 〈 0.05）. The main difference in the amounts of CH4 emission between the cultivars occurred in the period from the tillering stage to the heading stage. The biomass-scaled and yield-scaled CH4 emissions were respectively 3.8 and 5.2 mg/g for Ningjing 1, significantly lower than those for Zhendao 11 （7.4 and 12.8 mg/g, respectively）. According to the relationships between the plant growth characteristics and the CH4 emission, a stronger root system contributed mainly to the lower CH4 emission of Ningjing 1, as compared with Zhendao 11. Our results demonstrated that super rice has advantages not only in grain productivity but also in CH4 emission mitigation. Further expansion of super rice cropping will enhance rice yield and reduce greenhouse gas emission in China.     

Varietal Differences in Cell Wall β-(1→3),(1→4)-Glucan and Nonstructural Carbohydrate in Rice Stems during the Grain Filling Stage

Abstract

The contribution of cell wall components and nonstructural carbohydrate (NSC) to grain filling in rice (Oryza sativa L.) was clarified by investigating the differences in the dynamics of hemicellulose, sugar composition of hemicellulose, β-(1→3),(1→4)-glucan, and NSC among cultivars with different grain filling capacities. This investigation was performed using the stems of standard, high yield and low harvest index (HI) cultivars. Hemicellulose concentration in stems tended to decrease slightly during the grain filling stage. This decrease was attributed to a decrease in β-(1→3),(1→4)-glucan concentration, which was detected as a decrease in glucose composition of hemicellulose in the stems during the grain filling stage. The rate of decrease and decrease in the amount of β-(1→3),(1→4)-glucan in the stems differed among the cultivars. These were higher in high yield and high HI cultivars than in relatively low yield and low HI cultivars. Moreover, a positive correlation was observed between the rate of decrease in β-(1→3),(1→4)-glucan and NSC, indicating similarities in the dynamics of β-(1→3),(1→4)-glucan and NSC among the cultivars. When the top half of panicle was removed, β-(1→3),(1→4)-glucan and NSC concentrations in the culm and leaf sheath did not decrease during the grain filling stage. Therefore, the β-(1→3),(1→4)-glucan in stems might be one of the sources that supply substrate to panicle as well as NSC.     

Cold Tolerance of Core Collection at Booting Stage Associated with Eco-geographic Distribution in Yunnan Rice Landrace (Oryza sativa), China

Four hundred and seventy-seven accessions of the second core collection of rice landrace from five rice cropping regions (16 prefectures) of Yunnan Province, China, were evaluated for cold tolerance at booting stage. The results showed as follows: 1) there was a significant positive correlation (0.588**) between anther length and seed setting rate under natural low temperature conditions but was not significant in greenhouse (0.080). 2) there were significant differences in cold tolerance among core collections from different cropping regions under natural low temperature conditions while a negligible cold damage in greenhouse (with a seed setting rate approaching or exceeding 70%) for cold tolerance evaluation. Cold tolerance of core collection from Northwest Yunnan cold highland japonica region was the strongest, and that from south marginal paddy-upland rice region was the most sensitive, suggesting that the breeding goals for cold tolerance should be different in various rice cropping regions. 3) there were remarkable differences in cold tolerance of core collections from different prefectures of Yunnan Province under natural low temperature conditions. Based on the reduction of seed setting rate and characteristics of natural climate, the 16 prefectures could be divided into three categories, i.e. serious cold damage , cold damage and slight cold damage regions. 4) Difference of cold tolerance between different rice cropping regions and prefectures revealed further that temperature change caused by elevation and latitude was not only a dominant factor for differentiation of japonica and indica but also the basic reason that the genetic diversity and six ecological group of indica and japonica were being developed and, even the critical factor leading to the formation of the cold tolerance gene as well. The cold tolerance at booting stage could be believed to be resulted from the long term co-evolution between Yunnan rice landrce and cold stress in rice cropping     

Cadmium Accumulation and Its Toxicity in Brittle Culm 1(bc1),a Fragile Rice Mutant

Cadmium (Cd) accumulation and toxicity in rice plants were characterized and identified by using brittle culm 1 (bc1), a fragile rice mutant and its wild type (Shuangkezao, an indica rice) as materials by hydroponics. The low Cd level didn't obviously affect the growth parameters in both rice genotypes, but under high Cd levels (1.0 and 5.0 μmol/L), the growth of both rice plants were substantially inhibited. Moreover, bc1 tended to suffer more seriously from Cd toxicity than Shuangkezao. Cd accumulation in both rice plants increased with the increase of Cd levels. There was a significant difference in Cd accumulation between the two rice genotypes with constantly higher Cd concentration in bc1, which also accumulated more Cd at 0, 0.1, and 1.0 μmol/L Cd levels. The same case was found in the two rice plants grown on Cd-contaminated soil. This suggested that cell wall might play an important role in Cd accumulation in rice plants by the physiological mechanisms. The malondialdehyde (MDA) content, superoxide dismutase (SOD) and peroxidase (POD) activities in rice plants were affected differently under Cd treatments, and which implied that POD might play the main role in detoxifying active oxygen free radical. A significant difference in antioxidative system between the two rice genotypes was found with constantly higher MDA content, SOD and POD activities in bc1. In summary, bc1 accumulated more Cd and appeared to be more sensitive to Cd stress compared with its wild type.     

Analysis of Introgressed Segments in Near-isogenic Lines for F1 Pollen Sterility in Rice （Oryza sativa）

One hundred and fifty-eight microsatellite markers showing polymorphism among parents were used to survey the introgressed segments in the 50 near-isogenic lines of F1 pollen sterility. Two hundred and sixty introgressed segments were detected in 50 near-isogenic lines, each carrying 5.2 introgressed segments on an average. Among the 260 segments, one hundred carrying F1 pollen sterility loci concentrated on the region of F1 pollen sterility genes, and the remaining one hundred and sixty without F1 pollen sterility loci distributed randomly over 12 chromosomes.Both the average number and length of the introgressed segments decreased along with the increase of backcross generations. The number of introgressed segments was less than four and the length was less than 20 cM in the near-isogenic lines after backcrossing for four or more times.     

Tagging Blast Resistance Gene Pi 1 in Rice (Oryza sativa) Using Candidate Resistance Genes

An F3 population derived from C101LAC/CO39 containing 90 lines was analyzed for blast resistance with 48 candidate genes developed from resistance gene analogs (RGA) and suppression subtractive library. Genetic analysis confirmed that blast resistance of the population was controlled by a single gene Pi 1. One of the candidate genes, R10 was identified as associated with the blast resistance gene on the long arm of chromosome 11 and mapped using a DH population derived from Azucena/IR64. A pair of PCR based primers was designed based on the sequence of R10 for marker-aided selection of the blast resistance gene. The recombination frequency between Pi 1 and the marker was estimated as 1.28%. It suggested that strategy of employing candidate genes is useful for gene identification and mapping. A new RFLP marker and the corresponding PCR marker for tagging of Pi 1 were provided.     

Comparison of Grain Quality Characteristics Between F1 Hybrids and Their Parents in Indica Hybrid Rice

Sixteen widespread elite indica parents including seven CMS lines and nine restorer lines, and their 63 F1 hybrids were chosen to compare the eleven traits of grain quality. Overall results showed that the frequency of negative over-dominance, dominance and partial dominance was much higher than that of positive ones and heterosis, indicating that the values of F1 hybrids were generally lower than the means of their parents in quality characteristics. There existed apparent disparity in grain quality performance among F1 hybrids varied with the traits. The characteristics of chalky area percentage, chalky grain percentage, gel consistency and head rice recovery had a great variation in all kinds of tested heterosis indices including the mid-parent heterosis index, over high-value parent heterosis index and over low-value parent heterosis index, which suggested that special attention should be paid to the selection of F1 hybrids in these traits in breeding. The mean values of F1 hybrids were significantly lower than those of their higher parents in all the traits but close to or significantly lower than those of their mid-parent values except for kernel length and amylose content; and significantly higher than those of their lower parents except for brown rice recovery, milled rice recovery and gel consistency.     

Comparison of Grain Quality Characteristics Between F1 Hybrids and Their Parents in Indica Hybrid Rice

Sixteen widespread elite indica parents including seven CMS lines and nine restorer lines, and their 63 F1 hybrids were chosen to compare the eleven traits of grain quality. Overall results showed that the frequency of negative over dominance, dominance and partial dominance was much higher than that of positive ones and heterosis, indicating that the values of F1 hybrids were generally lower than the means of their parents in quality characteristics. There existed apparent disparity in grain quality performance among F1 hybrids varied with the traits. The characteristics of chalky area percentage, chalky grain percentage, gel consistency and head rice recovery had a great variation in all kinds of tested heterosis indices including the mid parent heterosis index, over high value parent heterosis index and over low value parent heterosis index, which suggested that special attention should be paid to the selection of F1 hybrids in these traits in breeding. The mean values of F1 hybrids were significantly lower than those of their higher parents in all the traits but close to or significantly lower than those of their mid parent values except for kernel length and amylose content; and significantly higher than those of their lower parents except for brown rice recovery, milled rice recovery and gel consistency.     

Analysis of Introgressed Segments in Near isogenic Lines for F1 Pollen Sterility in Rice (Oryza sativa)

One hundred and fifty eight microsatellite markers showing polymorphism among parents were used to survey the introgressed segments in the 50 near isogenic lines of F1 pollen sterility. Two hundred and sixty introgressed segments were detected in 50 near isogenic lines, each carrying 5.2 introgressed segments on an average. Among the 260 segments, one hundred carrying F1 pollen sterility loci concentrated on the region of F1 pollen sterility genes, and the remaining one hundred and sixty without F1 pollen sterility loci distributed randomly over 12 chromosomes. Both the average number and length of the introgressed segments decreased along with the increase of backcross generations. The number of introgressed segments was less than four and the length was less than 20 cM in the near isogenic lines after backcrossing for four or more times.