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Comparison of a chemical and enzymatic extraction of arsenic from rice and an assessment of the arsenic absorption from contaminated water by cooked rice.

Rice is a target food for arsenic speciation based analyses because of its relatively high arsenic concentration and per capita consumption rates. Improved speciation data for rice can be helpful in estimating inorganic arsenic exposures in the U.S. and in endemic populations. The inorganic arsenic exposure for cooked rice should include both the arsenic in raw rice plus the arsenic absorbed from the water used to prepare it. The amount of arsenic absorbed from water by rice during preparation was assessed using five different types of rice cooked in both contaminated drinking water and arsenic-free reagent water. The rice samples were extracted using trifluoroacetic acid (TFA) and speciated using IC-ICP-MS. The TFA procedure was able to extract 84-104% of the arsenic (As) from the five different cooked rice samples. Chromatographic recoveries ranged from 99% to 116%. The dimethylarsinic acid (DMA) and inorganic arsenic concentration ranged from 22 to 270 ng of As/g of rice and from 31 to 108 ng of As/g of rice, respectively, for samples cooked in reagent water. The overall recoveries, which relate the sum of the chromatographic species back to the total digested concentration, ranged from 89% to 117%. The absorption of arsenic by rice from the total volume of water [1:1 to 4:1 (water:rice)] used in cooking was between 89% and 105% for two different contaminated drinking water samples. A comparison of the TFA extraction to an enzymatic extraction was made using the five rice samples and NIST 1568a rice flour. The two extraction procedures produced good agreement for inorganic arsenic, DMA, and the overall recovery. Through the use of IC-ESI-MS/ MS with a parent ion of m/z 153 and fragment ions of m/z 138, 123, and 105, the structure dimethylthioarsinic acid was tentatively identified in two of the rice samples using the enzymatic extraction.

Absorption↗

Wild rice, hypoallergenic rice--immunologic comparison.

Rice is a cereal that is mainly produced and widely consumed in Asian countries including Korea. Several reports have suggested a role of IgE-mediated hypersensitivity in asthma and eczema associated with ingestion or inhalation of rice. In Japan, hypoallergenic rices are used for a substitute of common rice in some atopic patients. We performed this study to identify major allergens of rice and changed allergenicity in cooked and hypoallergenic rice. We purified crude extracts from a variety of rice and analyzed their protein distributions by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Based on UniCAP test and skin-prick test, we selected sera with high sensitivity and analyzed specific IgE binding to rice by immunoblotting. In addition, the inhibition rate among some rice was determined by enzyme linked immunosorbent assay and CAP test. As the result of this study, rice with various origins and polishing levels had no difference in protein band pattern. After cooking, it was difficult to detect protein bands distributed in raw rice; and, even through IgE immunoblot analysis, it was impossible to differentiate between wild and hypoallergenic rice. In addition, both wild and hypoallergenic rice still had IgE binding activity on their remaining protein bands. In conclusion, almost all proteins of rice were excluded or weakened in the process of boiling and IgE binding activity still remained even in hypoallergenic rice.

Antigens, Plant↗

Determination of selenium concentration of rice in china and effect of fertilization of selenite and selenate on selenium content of rice.

A method of hydride generation atomic fluorescence spectrometry was applied to the determination of the selenium concentration of regular polished rice in China and selenium-enriched polished rice obtained by foliar application of selenium-enriched fertilizer in the forms of selenite and selenate. The average selenium content of regular rice was 0.025 +/- 0.011 microg g(-)(1). On the basis of a daily dietary rice intake of 300-500 g suggested by the China Nutrition Society, the total selenium intake from regular rice was calculated to be 7.5-12.5 microg per person per day for an adult. The selenium contents of rice were significantly increased to 0.471-0.640 microg g(-)(1) by foliar application of selenium-enriched fertilizer at rate of 20 g of Se ha(-)(1) in the forms of sodium selenite and sodium selenate. The selenium content of rice by application of a fertilizer of selenate was 35.9% higher than that by a fertilizer of selenite, which showed that Se-enriched fertilizer in selenate exhibited greater efficiency in increasing Se content in rice products. The Se-enriched rice products can increase daily Se intake on average by 100-200 microg of Se per day by the consumption of 400 g of rice products if the Se level of rice products is controlled at 0.3-0.5 microg of Se g(-)(1). Because rice is a staple food in China, selenium-enriched rice obtained by bioenrichment of selenium to increase the Se content of rice could be a good selenium source for the population in selenium-deficient regions.

China↗

Protective effect of colored rice over white rice on Fenton reaction-based renal lipid peroxidation in rats.

Rice has been one of the most important grains. While polished white rice is favored, colored strains of rice, red, or black, have been maintained for religious purposes in Japan. We studied whether feeding of unpolished colored rice instead of white rice ameliorates oxidative renal tubular damage in rats induced by ferric nitrilotriacetate. Whereas renal lipid peroxidation was exacerbated in white rice-fed group in comparison with standard chow group, this exacerbation was not observed in red or black rice-fed groups. These changes were dependent on the proportion of colored rice to standard chow in the diet. Cyanidin 3-O-beta-D-glucoside was detectable neither in the serum nor kidney after one week of colored rice diet, but serum protocatechuic acid was significantly increased after black rice diet. There was a generalized decrease in the renal glutathione peroxidase activity in rice diet groups. Renal enzymatic activities of superoxide dismutase, glutathione S-transferase and NAD(P)H quinone reductase were not associated with the levels of lipid peroxidation. However, renal catalase activity was significantly increased in black rice-fed groups. These may partly explain the antioxidative effect. Furthermore, colored strains of rice are rich in proteins. Thus, our data warrants further investigation of the antioxidative effect of colored rice.

8-Hydroxy-2'-Deoxyguanosine↗

Novel euryarchaeotal lineages detected on rice roots and in the anoxic bulk soil of flooded rice microcosms

Because excised, washed roots of rice (Oryza sativa) immediately produce CH4 when they are incubated under anoxic conditions (P. Frenzel and U. Bosse, FEMS Microbiol. Ecol. 21:25-36, 1996), we employed a culture-independent molecular approach to identify the methanogenic microbial community present on roots of rice plants. Archaeal small-subunit rRNA-encoding genes were amplified directly from total root DNA by PCR and then cloned. Thirty-two archaeal rice root (ARR) gene clones were randomly selected, and the amplified primary structures of ca. 750 nucleotide sequence positions were compared. Only 10 of the environmental sequences were affiliated with known methanogens; 5 were affiliated with Methanosarcina spp., and 5 were affiliated with Methanobacterium spp. The remaining 22 ARR gene clones formed four distinct lineages (rice clusters I through IV) which were not closely related to any known cultured member of the Archaea. Rice clusters I and II formed distinct clades within the phylogenetic radiation of the orders "Methanosarcinales" and Methanomicrobiales. Rice cluster I was novel, and rice cluster II was closely affiliated with environmental sequences obtained from bog peat in northern England. Rice cluster III occurred on the same branch as Thermoplasma acidophilum and marine group II but was only distantly related to these taxa. Rice cluster IV was a deep-branching crenarchaeotal assemblage that was closely related to clone pGrfC26, an environmental sequence recovered from a temperate marsh environment. The use of a domain-specific oligonucleotide probe in a fluorescent in situ hybridization analysis revealed that viable members of the Archaea were present on the surfaces of rice roots. In addition, we describe a novel euryarchaeotal main line of descent, designated rice cluster V, which was detected in anoxic rice paddy soil. These results indicate that there is an astonishing richness of archaeal diversity present on rice roots and in the surrounding paddy soil.

Journal Article↗

Succession of methanogenic archaea in rice straw incorporated into a Japanese rice field: estimation by PCR-DGGE and sequence analyses.

The succession and phylogenetic profiles of methanogenic archaeal communities associated with rice straw decomposition in rice-field soil were studied by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis followed by 16S rDNA sequencing. Nylon bags containing either leaf sheaths or blades were buried in the plowed layer of a Japanese rice field under drained conditions during the off-crop season and under flooded conditions after transplanting. In addition, rice straw samples that had been buried in the rice field under drained conditions during the off-crop season were temporarily removed during spring plowing and then re-buried in the same rice field under flooded conditions at transplanting. Populations of methanogenic archaea were examined by amplification of the 16S rRNA genes in the DNA extracted from the rice straw samples. No PCR product was produced for samples of leaf sheath or blade prior to burial or after burial under drained conditions, indicating that the methanogen population was very small during decomposition of rice straw under oxic conditions. Many common bands were observed in rice straw samples of leaf sheath and blade during decomposition of rice straw under flooded conditions. Cluster analysis based on DGGE patterns divided methanogenic archaeal communities into two groups before and after the mid-season drainage. Sequence analysis of DGGE bands that were commonly present were closely related to Methanomicrobiales and Rice cluster I. Methanomicrobiales, Rice cluster I and Methanosarcinales were major members before the mid-season drainage, whereas the DGGE bands that characterized methanogenic archaeal communities after the mid-season drainage were closely related to Methanomicrobiales. These results indicate that mid-season drainage affected the methanogenic archaeal communities irrespective of their location on rice straw (sheath and blade) and the previous history of decomposition during the off-crop season.

Archaea↗

[Comparing study on oviposition preference and host-selecting behavior of larvae of rice and water-oat population or rice stem borer, Chilo suppressulis].

The oviposition preference of female of rice and water-oat population of rice stem borer was studied in the oviposition cage (1 m x 1 m x 0.8 m) in laboratory. The results showed that there was no significant difference between the numbers and distributions of eggs laid by the two populations, but each population preferred to lay its non-hatching ovum on the non-host plant. The host-selection behavior of larvae of two host populations was also investigated by Y-tube olfactory. When the leaf or leaf shell of rice and water-oat was provided to larvae of rice population in choice test, 4th and 6th instar larvae of rice population preferred rice leaf to water-oat leaf and 2nd and 4th instar larvae preferred leaf shell of rice to leaf shell of water-oat. When leaves of rice and water-oat were provided to larvae of water-oat population in choice test, except for 1st instar, each stadium of larvae preferred water-oat leaf to rice leaf, and when leaf shells of rice and water-oat were provided, all stadiums of larvae preferred leaf shell of water-oat to leaf shell of rice. These results demonstrated that intraspecies differentiation between the two populations, and the specialization of water-oat population of rice stem borer to host was higher than that of rice population.

Animals↗

Critical Importance of alpha-glucosidase contained in rice kernel for alcohol fermentation of rice polish.

In our previous study, we proposed a new alcoholic beverage called nuka-sake, which is made from uncooked rice polish without any enzyme source such as koji. In nuka-sake brewing, the uncooked rice polish serves not only as the fermentation material but also as the enzyme source. In the present study, the results of both laboratory-scale nuka-sake brewing runs with various grades of rice polish and analysis of amylolitic enzyme distribution in rice polish suggested that alpha-glucosidase (EC 3.2.1.20) is a key enzyme of parallel fermentation in nuka-sake brewing. Miglitol, a specific inhibitor of alpha-glucosidase, strongly inhibited glucose production from rice polish. To obtain further evidence regarding the contribution of alpha-glucosidase, this enzyme was purified from rice grain kernels (Yamadanishiki cultivar) and supplied for both rice polish saccharification and nuka-sake brewing. The purified alpha-glucosidase promoted both glucose production from rice polish and alcohol fermentation in nuka-sake brewing. Based on these results, it was considered that the alpha-glucosidase contained in rice polish plays an important role in glucose production. This role may be a rate-limiting factor for parallel fermentation in nuka-sake brewing. Moreover, oligosaccarides accumulated during the saccharification of uncooked rice polish, implying the contribution of not only alpha-glucosidase but also alpha-amylase (EC 3.2.1.1). Through this result, it can be speculated that the starch contained in rice polish will be decomposed to glucose as a result of the action of alpha-glucosidase and alpha-amylase contained in rice polish.

Journal Article↗

[Endosperm-specific expression of the ferritin gene in transgenic rice (Oryza sativa L.) results in increased iron content of milling rice].

Iron deficiency is the most widespread micronutrient deficiency world-wild, and is estimated to affect about 30% of the world population. To increase the iron content of rice in Chinese, the 764 bp cDNA of ferritin gene was cloned from soybean (Phaseolus limensis), and constructed between the 1 353 bp rice glutelin GluB-1 promoter and NOS terminator in a binary vector pCAMIBA1301. The constructed pYF1067 vector was introduced into Agrobacterium strain EHA105, and used for transformation of the primary callus derived from immature embryos of a high-yielding rice ( Oryza sativa L. ssp. japonica) variety Wuxiangjin 9. Under the selection on hygromycin-containing medium, seventeen independent transgenic rice lines, total more than 80 transgenic plants, were finally regenerated, and most of these transgenic rice plants grew normally. PCR and Southern blot analysis of total DNA from primary transformants confirmed that one to three copies of the transgenes integrated into the genome of most of the transgenic plants, and they could be stably transmitted into the progeny of the transgenic rice. Northern blot analysis showed that the ferritin gene could specifically express in the endosperm of transgenic rice with high level, while no or low expression in leaves. The expression level varied among different independent transgenic rice plants. There was a significant effect of the high-expression of ferritin on the increased iron content in transgenic rice, the iron content in the milling rice of transgenic rice was up to 64% higher than that of the untransformed wild-type plant, whereas no significant alteration of the zinc level occurred between these two type rice plants.

Ferritins↗

[Specific expression of the foreign gene regulated by the rice rbcS promoter in transgenic rice].

To use different types of promoters in transgenic rice research, the 5'-upstream regulation region of rice Rubisco small subunit gene (rbcS) was cloned from a Chinese cultivar Wuyunjing 8, and its sequences were confirmed by comparison with the known genome sequences of both japonica and indica rice. The cloned rbcS promoter was fused to the 5'-upstream of GUS (beta-glucuronidase) coding region in a binary vector, and introduced into rice by Agrogacterium-mediated transformation. The integration of the rbcS-GUS fusion gene in transgenic rice was confirmed by PCR analysis. The results of both histochemical staining and quantitative analysis of GUS activity showed that the expression level of GUS fusion gene was significantly stronger in leaf blade and sheath than in other organs of transgenic rice plants, and the GUS activity was restricted to the mesophyll cells of leaf tissue, which showed that the rice rbcS promoter could control not only the tissue- but also the cell-specific expression of foreign genes in transgenic rice. The present results also demonstrated that light induction had a significant effect on the enhancement of transgene's expression when regulated by the rice rbcS promoter in transgenic rice. Our results showed that the rice rbcS promoter might be very useful for the expression of target genes in transgenic rice, with particularly high efficiency in leaf tissues.

Gene Expression Regulation, Plant↗

Genetic differentiation for nuclear, mitochondrial and chloroplast genomes in common wild rice ( Oryza rufipogon Griff.) and cultivated rice ( Oryza sativa L.).

The genetic differentiation of nuclear, mitochondrial (mt) and chloroplast (cp) genomes was investigated by Southern and PCR analysis using 75 varieties of cultivated rice ( Oryza sativa L.) and 118 strains of common wild rice (CWR, Oryza rufipogon Griff.) from ten countries of Asia. The distinguishing differences between the Indica and Japonica cultivars were detected both in the nuclear genome and the cytoplasmic genome, confirming that the Indica-Japonica differentiation is of major importance for the three different classes of genome in cultivated rice. This differentiation was also detected in common wild rice with some differences among the genome compartments and the various regions. For nuclear DNA variation, both Indica-like and Japonica-like types were observed in the Chinese CWR, with the latter more-frequent than the former. No Japonica-like type was found in South Asia, and only two strains of the Japonica-like type were detected in Southeast Asia, thus the Indica-like type is the major type among South and Southeast Asian CWR. For mtDNA, only a few strains of the Japonica-like type were detected in CWR. For cpDNA, the Japonica type was predominant among the CWR strains from China, Bangladesh and Burma, while the Indica type was predominant among the CWR strains from Thailand, Malaysia, Cambodia and Sri Lanka, and both types were found in similar frequencies among the Indian CWR. Altogether, however, the degree of Indica-Japonica differentiation in common wild rice was much-less important than that in cultivated rice. Cluster analyses for nuclear and mitochondrial DNA variation revealed that some CWR strains showed large genetic distances from cultivated rice and formed clusters distinct from cultivated rice. Coincidence in the genetic differentiation between the three different classes of genome was much higher in cultivated rice than in CWR. Among the 75 cultivars, about 3/4 entries were "homoeotype" showing congruent results for nuclear, mt and cpDNA regarding the Indica-Japonica differentiation. In CWR, the proportions of homoeotypes were 5.7%, 15% and 48.8% in China, South Asia and Southeast Asia, respectively. Based on the average genetic distance among all the strains of CWR and cultivated rice for nuclear and mitochondrial genomes, the variability of the nuclear genome was found to be higher than that of the mitochondrial genome. The global pattern based on all genomes shows much-more diversification in CWR than that in cultivated rice.

Journal Article↗

Light-regulated and cell-specific expression of tomato rbcS-gusA and rice rbcS-gusA fusion genes in transgenic rice.

A previously isolated rice (Oryza sativa) rbcS gene was further characterized. This analysis revealed specific sequences in the 5' regulatory region of the rice rbcS gene that are conserved in rbcS genes of other monocotyledonous species. In transgenic rice plants, we examined the expression of the beta-glucuronidase (gusA) reporter gene directed by the 2.8-kb promoter region of the rice rbcS gene. To examine differences in the regulation of monocotyledonous and dicotyledonous rbcS promoters, the activity of a tomato rbcS promoter was also investigated in transgenic rice plants. Our results indicated that both rice and tomato rbcS promoters confer mesophyll-specific expression of the gusA reporter gene in transgenic rice plants and that this expression is induced by light. However, the expression level of the rice rbcS-gusA gene was higher than that of the tomato rbcS-gusA gene, suggesting the presence of quantitative differences in the activity of these particular monocotyledonous and dicotyledonous rbcS promoters in transgenic rice. Histochemical analysis of rbcS-gusA gene expression showed that the observed light induction was only found in mesophyll cells. Furthermore, it was demonstrated that the light regulation of rice rbcS-gusA gene expression was primarily at the level of mRNA accumulation. We show that the rice rbcS gene promoter should be useful for expression of agronomically important genes for genetic engineering of monocotyledonous species.

Amino Acid Sequence↗

Survival of Azorhizobium caulinodans in the Soil and Rhizosphere of Wetland Rice under Sesbania rostrata-Rice Rotation.

The survival of indigenous and introduced strains of Azorhizobium caulinodans in flooded soil and in the rice rhizosphere, where in situ Sesbania rostrata was incorporated before the rice crop, is reported. The azorhizobia studied were both root and stem nodulating. In a pot experiment, two crop cycles each of inoculated and noninoculated Sesbania-rice were compared with two crop cycles of flooded fallow-rice. In a field experiment, the effect of repeated incorporation of in situ S. rostrata in the Sesbania-rice sequence was studied. Soils in which inoculated S. rostrata was incorporated contained about 3,000 times more azorhizobia than did soils in the flooded fallow treatment and about 50 times more azorhizobia than did soils in the noninoculated Sesbania treatment. Azorhizobial numbers in the inoculated Sesbania treatment declined toward rice harvest but remained much higher than in the flooded fallow-rice treatment. Repeated incorporation of S. rostrata increased the population density of indigenous soil azorhizobia, whereas the population of inoculated strain ORS571 (Str Spc) declined to an undetectable level; this finding suggested low competitiveness by the introduced strain. In the incorporated Sesbania treatment, the rice rhizosphere harbored significantly more A. caulinodans and supported higher nitrogenase activity per plant than did the rhizosphere of the flooded fallow-rice treatment. Sterile rice seedlings inoculated with A. caulinodans showed nitrogenase activity comparable to that of seedlings inoculated with Azospirillum lipoferum 34H, a rice root isolate. Rhizobia from Sesbania aculeata, Sesbania sesban, a Trifolium sp., and Vigna unguiculata did not support appreciable nitrogenase activity.

Journal Article↗

Growth of nisin-producing lactococci in cooked rice supplemented with soybean extract and its application to inhibition of Bacillus subtilis in rice miso.

Lactic acid fermentation of cooked rice and rice koji by supplementation with soybean extract (SBE) and its application to rice miso fermentation were investigated. By supplementing the cooked rice with SBE, lactic acid bacteria (LAB) grew well without any unfavorable effects on the rice such as off-flavor or coloration. Lactococcus lactis subsp. lactis IFO12007 (Lc. lactis, a producer of the bacteriocin nisin) proliferated at 10(8 to approximately 9) cells/g after 24 h of incubation and produced high activity of nisin. The fermented rice with Lc. lactis strongly inhibited not only Bacillus subtilis ATCC19659 but also the other Bacillus strains. While some strains of LAB markedly inhibited the growth of Asp. oryzae, resulting in failure of koji fermentation, Lc. lactis did not affect the growth of these molds. When Lc. lactis was used for rice miso fermentation as a lactic acid starter culture, Lc. lactis rapidly proliferated and produced high nisin activity of 6,400 IU/g, in the steamed rice, resulting in complete growth inhibition of B. subtilis, which had been inoculated at the beginning of the koji fermentation. The rice miso after 12 weeks of aging had a suitable pH, and favorable taste and color. Furthermore, hyposalting of rice miso could be done without difficulty by lactic acid fermentation of both rice and soybeans.

Bacillus subtilis↗

[Construction of chromosome segment substitution lines carrying overlapping chromosome segments of the whole wild rice genome and identification of quantitative trait loci for rice quality].

Rice is one of the major staple cereal grains. Most of the important traits of crops are complex traits controlled by quantitative trait loci (QTL). Detection and genetic identification of QTLs can provide insights into molecular and biological mechanisms of development and physiology. Chromosome segment substitution lines (CSSLs) have been proposed as a simple and powerful way to identify QTLs. The demand for rice grain quality becomes increasingly important. The primary components of rice grain quality include appearance, milling and physico-chemical, cooking and eating quality. Most of these traits are complex and controlled by QTLs, so genetic characterization of these traits is more difficult than that of traits each controlled by a singular gene such as Waxy, which controls glutinousness of rice grain. We constructed 133 chromosome segment substitution lines (CSSLs) from backcross progenies (BC3F2) derived from a cross between Teqing (an O. sativa L. ssp. indica variety) as the recurrent parent and wild rice (O. rufipogon Griff.) as the donor parent. In this process, we carried out marker-assisted selection (MAS) by using 118 CAPS or SSR markers covering the whole rice genome (Figs.1, 2). Because phenotypic values of quality trait were significantly different between cultivar Teqing and wild rice, the CSSLs were used to identify QTLs for rice quality traits. Three appearance quality traits (head rice percentage, HRP; percentage of chalky rice grains, PCRG; transparency, TP) and five physico-chemical properties (gelatinization temperature, GT; gel consistency, GC; amylose content, AC; protein content, PC; fat content, FC) were scored using 133 CSSLs which substituted segments covering the whole genome of wild rice. A total of 15 QTLs for five of the eight traits mentioned above (HRP, PCRG, TP, PC and FC) were identified on nine chromosomes, and several QTLs affecting different quality traits were mapped in the same regions (Fig.3, Table 1).

Chromosomes, Plant↗

Effect of two rice culture methods on the seasonal occurrence of mosquito larvae and other aquatic animals in rice fields of southwestern Korea.

An ecological study has been performed on the community structures of aquatic animals and on abiotic factors in organically and conventionally-farmed rice fields at Bulkyo, Bosong-gun, Chollanamdo during the rice growing periods between May 1995 and October 1996. The total numbers of aquatic insect taxa in these fields were 25 species, 22 families in 10 orders. In 1995 and 1996, three and then four more species of aquatic insects were collected in the organically-farmed rice fields than in conventionally-farmed rice fields. The common dominant species in both rice fields during the rice cultivation period was Chironomus sp. The abundance of two vector mosquitoes, Anopheles sinensis and Culex tritaeniorhynchus, was lower in the organically-farmed rice fields as compared to the conventionally-farmed rice fields. The populations of mosquito larvae in the organically-farmed rice fields might have been influenced by Chinese muddy loaches, Misgurnus mizolepis. Coefficients of correlation between Chinese muddy loaches and abundance of mosquito larvae showed negative correlations in An. sinensis (-0.66) and Cx. tritaeniorhynchus (-0.47). The average of the species diversity index for organically-farmed rice fields was almost twice (0.62) as much as is of conventionally-farmed rice fields (0.35). The average community similarity between both types of fields was very low (0.33).

Agriculture↗

One large-insert plant-transformation-competent BIBAC library and three BAC libraries of Japonica rice for genome research in rice and other grasses.

We report one large-insert BIBAC library and three BAC libraries for japonica rice cv Nipponbare. The BIBAC library was constructed in the HindIII site of a plant-transformation-competent binary vector (pCLD04541) and the three BAC libraries were constructed in the BamHI, HindIII and EcoRI sites of a BAC vector (pECBAC1), respectively. Each library contains 23,040 clones, has an average insert size of 130 kb, 170 kb, 150 kb and 156 kb, and covers 6.7x, 8.7x, 7.7x and 8.0 x rice haploid genomes, respectively. The combined libraries contain 92,160 clones in total, covering 31.1 x rice haploid genomes. To demonstrate their utility, we screened the libraries with 55 DNA markers mapped to chromosome 8 of the rice genetic maps and analyzed a number of clones by the restriction fingerprinting and contig assembly method. The results indicate that the libraries completely cover the rice genome and, thus, are well-suited for genome research in rice and other gramineous crops. The BIBAC library represents the first plant-transformation-competent large-insert DNA library for rice, which will streamline map-based cloning, functional analysis of the rice genome sequence and molecular breeding in rice and other grass species. These libraries are being used in the development of a whole-genome, BAC/BIBAC-based, integrated physical, genetic and sequence map of rice and in the research of genome-wide comparative genomics of grass species.

Journal Article↗

Toxicity of copper on rice growth and accumulation of copper in rice grain in copper contaminated soil.

Pot soil experiments showed that copper (Cu) is highly toxic to rice. Rice grain yields decreased exponentially and significantly with the increase of soil Cu levels. Rice grain yield was reduced about 10% by soil Cu level of 100 mg kg(-1), about 50% by soil Cu level of 300-500 mg kg(-1) and about 90% by soil Cu concentration of 1,000 mg kg(-1). Root was more sensitive to soil Cu toxicity than other parts of rice plant at relatively lower soil Cu levels (less than 300-500 mg kg(-1)), but the growth of whole rice plant was severely inhibited at high soil Cu levels (300-500 mg kg(-1) or above). Cu concentrations in rice grain increased with soil Cu levels below 150-200 mg kg(-1), but decreased with soil Cu levels above 150-200 mg kg(-1), with peak Cu concentration at soil Cu level of 150-20 mg kg(-1). Cu was not distributed evenly in different parts of rice grain. Cu concentration in cortex (embryo) was more than 2-fold that in chaff and polished rice. More than 60% of the Cu in grain was accumulated in polished rice, about 24% in cortex (embryo), and about 12% in chaff. So, about 1/3 of the Cu in rice grain was eliminated after grain processing (chaff, cortex and embryo was removed).

Copper↗