Carbon flux from decomposing 13C-labeled transgenic and nontransgenic parental rice straw in paddy soil

Purpose

Genetic modification of Bt rice may affect straw decomposition and soil carbon pool under flood conditions. This study aims to assess the effects of cry gene transformation in rice on the residue decomposition and fate of C from residues under flooded conditions.

Materials and methods

A decomposition experiment was set up using ¹³C-enriched rice straws from transgenic and nontransgenic Bt rice to evaluate the soil C dynamics and CH₄ or CO₂ emission rates in the root and non-root zones. The concentrations and stable carbon isotope compositions of the soil organic carbon (SOC), dissolved organic carbon (DOC), microbial biomass carbon (MBC), CH₄, and CO₂ of the root and non-root zones were determined from 7 to 110 days after rice straw incorporation.

Results and discussion

Rice straw incorporation into soil significantly increased the SOC, DOC, and MBC concentrations and the CH₄ and CO₂ emission rates. The percentage of ¹³C-SOC remaining in the root zone was significantly lower than that in the non-root zone with rice straw decomposition. The DOC and MBC concentrations significantly increased in both the root and non-root zones between 0 and 80 days after rice straw incorporation. However, no significant differences were found after Bts (Bt rice straw added into soil) and Cks (nontransgenic Bt rice straw added into soil) incorporation in the root and non-root zones. This result may be attributed to the priming effects of sufficient oxygen and nutrients on straw degradation in the root zone.

Conclusions

Bt gene insertion did not affect the SOC, DOC, and MBC concentrations and the CH₄ and CO₂ emission rates in both the root and non-root zones. However, rice straw incorporation and root exudation significantly increased the SOC, DOC, and MBC concentrations and the CH₄ and CO₂ emission rates.     

Storage Stability of Flour‐Blasted Brown Rice

Brown rice was blasted with rice flour rather than sand in a sand blaster to make microperforations so that water could easily penetrate the brown rice endosperm and cook the rice in a shorter time. The flour‐blasted American Basmati brown rice, long‐grain brown rice, and parboiled long‐grain brown rice samples were stored in Ziploc storage bags under atmospheric conditions and in vacuum‐packed bags. They were periodically tested for over 10 months for changes in water absorption, free fatty acid (FFA), peroxide value (POV), viscosity changes of flour using the Rapid ViscoAnalyser (RVA), and texture of whole cooked kernel using a texture analyzer during cooking. Flour‐blasted brown rice absorbed less water but needed less cooking time than its counterpart that was not flour‐blasted. There was an increase in FFA, POV, peak viscosity (PV), final viscosity (FV), breakdown viscosity (BD), and setback viscosity (SB) during storage of flour‐blasted brown rice for 300 days, but no change was observed in texture (hardness, gumminess) and water absorption. The combined coefficient of correlation (including all types of rice) between FFA and FV is r = 0.86 and between FFA and SB is r = 0.90 at P < 0.0001.     

Transgenic Bt rice does not affect enzyme activities and microbial composition in the rhizosphere during crop development

Use of transgenic crops, including those expressing the insecticidal Cry protein from Bt, is increasing at a rapid rate in worldwide. Field and laboratory studies of transgenic Bt crops have been carried out to detect the persistence and activity of the Cry protein in soil and its effect on soil microorganisms to assess their risks to environment. However, there were few studies that evaluate the seasonal effects of Bt rice on rhizosphere soil microbial communities compared to those of insecticides commonly applied in paddy soil for the control of lepidopteran insects. In this study, seasonal effects of transgenic rice expressing the Cry1Ab insecticidal protein active against lepidoperan pests and the insecticide triazophos [3-(o,o-diethyl)-1-phenyl thiophosphoryl-1,2,4-triazol] on soil enzyme activities and microbial communities were compared under field conditions. During a 2-year field study, rhizosphere soil samples of transgenic-Bt rice (Bt), non-Bt parental rice (Ck) and non-Bt parental rice with triazophos (Ckp) applied were taken at four stages in the rice developmental cycle: seedling, booting, heading and maturing. Microbial processes were investigated by measuring different biochemical activities including those involved in C and P cycling. Denaturing gradient gel electrophoresis (DGGE) and terminal-restriction fragment length polymorphism (T-RFLP) analyses were used to compare rhizosphere microbial compositions. Some occasional and inconsistent effects of the application of triazophos on the bacterial composition in the rhizosphere soil of rice plant were found at the booting and heading stages as compared with that of transgenic-Bt rice. There were no statistically significant differences (P>0.05) in phosphatase activity, dehydrogenase activity, respiration, methanogenesis or fungal community composition in rhizosphere soil between Bt, Ck and Ckp over the rice cropping cycle. However, seasonal variations in the selected enzyme activities and microbial community composition in the rhizosphere soil of Bt, Ck and Ckp were clearly detected. These results suggested that the changes in rhizophere soil microbial community composition associated with the crop growth stage overweighed the application of triazophos and the cry1Ab gene transformation. KMD1 (Bt) rice expressing the cry1Ab gene had no measurable adverse effect on the key microbial processes or microbial community composition in rhizophere soil over 2 years of rice cropping.     

Transgenic Enhancement of High‐Molecular‐Weight Glutenin Subunit 1Dy10 Concentration: Effects in Wheat Flour Blends and Sponge and Dough Baking

Dough strength is needed for efficient breadmaking quality. This property is strongly influenced in wheat (Triticum aestivum L.) by gluten seed storage proteins and, in particular, by high‐molecular‐weight (HMW) glutenin subunit composition. Experiments were designed to elevate expression of a key native HMW glutenin subunit (1Dy10) via genetic engineering and to determine whether resultant flours can be used in sponge and dough applications, the most common commercial bread‐baking procedure. Both unblended and blended samples from transgenic and nontransgenic sister lines were tested, with blended samples being formed by addition to a control sample. Dough properties, as determined by farinograph evaluation, were improved by the transgene‐encoded increases in 1Dy10 in both undiluted and blended flours. Mean farinograph stability of transgenic samples was twice that of the control, and blends with transgenic samples demonstrated increases in stabilities proportional to the amount of transgenic flour included. Mean farinograph quality numbers of transgenic samples, and of all blends containing transgenic flour, were significantly higher than both the control and all nontransgenic treatments. In the sponge and dough bake procedure, undiluted transgenic samples induced lower scores, relative to both control and undiluted nontransgenic samples, for water absorption, crumb body firmness, and loaf volume. In blends, however, the transgenic samples resulted in improvements in some sponge and dough loaf attributes, including loaf symmetry and crumb color score, without any concomitant loss of loaf volume in transgenic blends. These improved variables relate to finished product appearance and to consumer selection in markets. The use of transgenic flours with increased 1Dy10 glutenin content in commercial blends could provide advantages in sponge and dough bake applications.     

Soil persistence of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> (Bt) toxin from transgenic Bt cotton tissues and its effect on soil enzyme activities

A silty loam soil was incubated with the leaves and stems of two transgenic Bacillus thuringiensis (Bt) cotton varieties and nontransgenic Bt cotton to study the soil persistence of the Bt toxin from the decomposing transgenic Bt cotton tissues and its effect on soil enzyme activities. The results showed that after Bt cotton tissue amendment, Bt toxin was introduced into soil upon decomposition; about 50% of the introduced Bt toxin persisted in soil for at least 56 days. No Bt toxin was detected in the nontransgenic Bt cotton-amended soil; the amount of Bt toxin was the highest in the soil treated with the residue with the higher Bt toxin content. Activities of soil urease, acid phosphomonoesterase, invertase, and cellulase were stimulated by the addition of Bt cotton tissues, whereas activity of soil arylsulfatase was inhibited. Probably cotton tissue stimulated microbial activity in soil, and as a consequence, enzyme activities of soil were generally increased. This effect can mask any negative effect of the Bt toxin on microbial activity and thus on enzyme activities.     

转Bt-Cry1Ac基因棉花对烟粉虱体内几种酶活力的影响

为研究转Bt基因棉花上烟粉虱种群较常规棉上升的原因是否受到Bt毒蛋白的影响,以及是否与体内保护酶和解毒酶活力变化有关,应用转Bt基因棉花‘新棉33B’及其常规棉受体‘33’为试验对象,采用ELISA法和酶活力测定法,分别研究了取食转Bt基因棉花后烟粉虱体内Bt毒蛋白含量以及体内过氧化氢酶(CAT)、过氧化物酶(POD)、超氧化物歧化酶(SOD)3种保护酶和乙酰胆碱酯酶(AChE)、羧酸酯酶(CarE)和谷胱甘肽-S-转移酶(GSTs)3种解毒酶活力的变化情况。结果表明,在取食转Bt基因棉花‘新棉33B’4 h后,烟粉虱体内能检测到Cry1Ac蛋白,并且在12 h后维持在一个相对稳定的状态。取食‘新棉33B’后烟粉虱体内的SOD和GSTs活力受到显著抑制(P0.05),并且随着取食时间的延长,SOD活力逐渐下降,其中取食8 h、12 h、24 h和36 h后较取食‘33’的对照分别下降了37.8%、32.1%、32.0%和31.9%。CAT、POD和CarE活力显著提高(P0.05),并且随着取食时间的延长,酶活力逐渐上升,与对照相比,取食12 h、24 h和36 h后,CAT活力分别为取食‘33’的对照的1.54倍、1.55倍和1.42倍;POD活力分别为取食‘33’的对照的1.59倍、1.39倍和1.53倍;CarE活力分别为取食‘33’的对照的1.32倍、1.34倍和1.39倍;取食‘新棉33B’对AChE活力没有明显影响。结果提示,转Bt-Cry1Ac基因棉花对烟粉虱保护酶活力总体起到促进作用,对解毒酶活力总体影响不大。故烟粉虱体内保护酶活力的增加可能会有益于烟粉虱种群的增长,但是否起到决定性作用还有待进一步研究确定。     

Bt水稻杀虫蛋白时空变化及秸秆还田后在土壤中的持留规律

以Bt抗虫水稻华池B6、TT51及其非转基因水稻亲本嘉早935、明恢63,以及与它们亲缘较远但农艺性状相近的水稻品种中九B、R9311为试验材料,研究了田间种植条件下Bt抗虫水稻杀虫蛋白的时空变化及其在根际土中的持留规律,同时,还研究了秸秆还田后Bt蛋白在土壤中的持留规律。结果表明：1）Bt抗虫水稻华池B6植株各个部位...     

Effect of Transgenic Bt Cotton on Bioactivities and Nutrients in Rhizosphere Soil

The influence of transgenic Bacillus thuringiensis (Bt) cotton (BtXincai1) and its corresponding nontransgenic isoline (Xincai1) on the microorganisms, enzyme activity, and nutrient content of rhizosphere soil was studied through experiments in potted plants. The calcareous drab soil samples were collected (0–15 cm deep) from an experimental field in Shanxi Agricultural University (China) in 2005. The pots were categorized in different groups with replicates for each variety (transgenic BtXincai1 and general Xincai1). The rhizosphere soil samples were collected at different growth periods (seedling, bud, flowering, peak boll, boll opening, and harvest). The Bt protein and other microbial properties in the soil samples were determined by using selected methods (material and methods session). The results demonstrated that the concentration of the Bt protein in the rhizosphere soil of BtXincai1 reached a peak at 56.14 ng g^?1 during the flowering period. However, the Bt protein would not continuously accumulate in the soil. The rhizosphere soil of BtXincai1 was more suitable for the growth and proliferation of bacteria and fungi but it had no significant impact on the number of actinomycetes. BtXincai1 had some inhibitory effects on alkaline phosphatase activity in the rhizosphere soil, and it might promote dehydrogenase activity during the blooming period. However, it had no significant influence on protease, urease, or sucrase activities. Further, it had no significant impact on the contents of organic matter, total nitrogen, available nitrogen, or potassium in rhizosphere soil. It could significantly decrease the content of available phosphorus during the flowering period. Based on this study, the sensitive reactions of microorganisms and the activities of alkaline phosphatase and dehydrogenase might be considered as three potential indexes for assessing the risk posed by transgenic Bt cotton to soil ecology.     

苏云金芽孢杆菌cry1Ia基因的克隆、表达与活性研究

根据cry1Ia类基因的全长序列设计引物，以苏云金芽孢杆菌（Bacillus thuringiensis）菌Btc008的总DNA为模板扩增出片段长为2.1kb的cry1Ia的全长基因，插入大肠杆菌（Escherichia coli）表达载体pET-21b，转化大肠杆菌BL21（DE3）菌株，诱导表达出81kD的蛋白。该蛋白由719个氨基酸组成，推导的分子量为81.2kDa。该蛋白的氨基酸序列不同于已知的12种Cry1Ia蛋白，是一种新的Cry1Ia蛋白，该基因已被国际基因命名委员会正式命名为cry1Ia8。杀虫活性测定结果表明：Cry1Ia8对亚洲玉米螟（Ostrinia furnacalis）、小菜蛾（Plutella xylostella）有很强的杀虫活性，LC50分别为0.268 µg/g、2.227 µg/ml，其杀虫效果与Cry1Ab、Cry1Ac相当。对大豆食心虫（Leguminivora glycinivorella）也有较好的活性，但对鞘翅目叶甲科害虫榆兰叶甲（Pyrrhalta aenescens）没有活性。该基因的获得将为我国抗虫转基因作物和工程菌的研制提供新的基因来源，为筛选延缓昆虫抗性产生的基因组合提供了极为重要的依据。     

Fate of Bt Protein and Influence of Corn Hybrid on Ethanol Production

Corn hybrids were compared to determine the fate of recombinant Bt protein (CRY1Ab from Bacillus thuringiensis) in coproducts from dry grind and wet‐milled corn during production of fuel ethanol. Two pairs of Bt and non‐Bt hybrids were wet milled, and each fraction was examined for the presence of the Bt protein. Bt protein was found in the germ, gluten, and fiber fractions of Bt hybrids. In addition, one set of Bt and non‐Bt hybrids were treated by the dry‐grind ethanol process and Bt protein was monitored during each step of the process. The Bt protein was not detected after liquefaction. Subsequent experiments determined that the Bt protein is rapidly denatured at liquefaction temperatures. Finally, five hybrids were compared for ethanol yield after dry grinding. Analysis of fermentation data with an F‐test revealed the percent of total starch available for conversion into ethanol varied significantly among the hybrids (P < 0.002), indicating ethanol yield is not exclusively dependent on starch content. No difference, however, was observed between Bt and non‐Bt corn hybrids for either ethanol productivity or yield.     

苏云金芽孢杆菌cry2Aa基因的克隆、表达与活性

B-8-G和Ly30是我国自行分离的对多种重要农业害虫具有高毒力的苏云金芽孢杆菌（Bacillus thuringiensis）菌株,经PCR-RFLP鉴定均含有cry2Aa基因.根据cry2Aa全长基因序列设计特异引物,以B-8-G总DNA为模板扩增其中的cry2Aa全长基因,与大肠杆菌（Escherichia coli）表达载体pET-21b相连接,获得含有cry2Aa全长基因的重组质粒pET2Aa,该基因在大肠杆菌BL21菌株能够正常表达65 kD蛋白.通过构建Ly30总DNA文库方法从中筛选获得cry2Aa基因,将其连接至Bt-E.coli穿梭表达载体pHT315上,转化Bt无晶体突变株HD-73中,该基因能正常表达65 kD蛋白,并形成立方体状晶体.这两种基因序列已被国际Bt基因命名委员会分别正式命名为cry2Aa9和cry2Aa10.杀虫生物活性测定结果表明cry2Aa基因表达产物对黄胫小车蝗（Oedaleusinfernlis）、尖音库蚊（Culex pipiens）、黑翅伊蚊（Aedes melanopterus）、水稻二化螟（Chilo suppressalis）和小菜蛾（Plutellaxylostella）幼虫均具有显著的毒杀作用.首次报道cry2Aa10基因表达蛋白对蝗虫、库蚊具有杀虫活性.这些基因的获得,将为高效工程菌和抗虫转基因植物的研制提供了新的基因资源.     

Soil nematode community varies between rice cultivars but is not affected by transgenic Bt rice expressing <Emphasis Type="Italic">Cry1Ab</Emphasis> or <Emphasis Type="Italic">Cry1Ab</Emphasis>/<Emphasis Type="Italic">Cry1Ac</Emphasis>

Insecticidal crystal (Cry) proteins produced by transgenic Bacillus thuringiensis (Bt) rice that enter the soil via root exudation and plant residues may be harmful to non-target soil organisms. We conducted a 3-year field investigation to determine if soil nematode abundance and diversity were affected by exposure to two transgenic Bt rice cultivars, compared to their non-transgenic near isolines. Near isolines were Kemingdao (KMD-Bt) expressing the single Cry1Ab gene and its non-Bt near isoline Xiushui-11 (XSD), as well as Huahui-1 (HH-Bt) expressing the fused Cry1Ab/Cry1Ac gene and its non-Bt near isoline Minghui-63 (MH). Nematode variables including community composition, abundance, trophic groups, and most of the common genera differed significantly between the rice cultivars. However, these nematode variables were similar under transgenic Bt rice and its non-Bt near isoline, although higher Shannon’s index value and Pielou’s index value were found in soils planted with Bt rice than the non-Bt near isoline. During this 3-year field study, gene modification (single Cry1Ab gene and fused Cry1Ab/Cry1Ac gene) supports a more uniform distribution of nematode species but had no effect on soil nematode abundance and community composition. We conclude that continuous cultivation of KMD-Bt and HH-Bt rice varieties for 3 years is not detrimental to soil nematode communities under field conditions.     

内源性土壤苏云金杆菌(BRC-ZYR2)对含氧铬还原的影响研究

Chromium（Cr） may cause losses in the yield of field plant, which is one of the favorite habitats of Bacillus thuringiensis（Bt）. The purposes of our study were to assess the Cr（VI）-resistance and Cr（VI）-reducing abilities of an indigenous soil isolate of Bt and to determine the factors governing Cr（VI） reduction. Towards this end a novel dichromate-reducing Bt BRC-ZYR2, characterized with insecticidal crystal proteins（ICPs）, was isolated from a uranium deposit. Minimum inhibitory concentrations（MICs） of Cr（VI） were determined by broth dilution method and the concentrations of Cr（VI） and total Cr in the supernatant were quantified colorimetrically using 1,5-diphenylcarbazide（DPC） reagent and a mixture of sulfuric-nitric acids, respectively. The isolate contained five ICP genes（cry1Ba, cry1 Bb, cry1Be/cry1 Bf, cry9 Ca and cry9Da） and exhibited a high level of Cr（VI） resistance with MICs of 150 mg L-1at pH 7.0 and 30？C, and 500 mg L-1under optimal conditions（pH 9.0 and 40？C）. The total Cr concentration was similar to initial concentration of Cr（VI） under the optimal condition, suggesting that the essential removal of the Cr（VI） was dependent on Bt reduction. Under optimal conditions, the initial Cr（VI） concentrations from 25 to 75 mg L-1significantly decreased in 24 h after incubation. Addition of Mn2＋, Co2＋, Mo2＋and Cu2＋activated Bt-mediated Cr（VI） reduction, while Zn2＋, Ni2＋and glucose were found to inhibit the reduction. Our results indicated that this isolate could be a promising biopesticide with the potential for both insect biocontrol and Cr bioremediation in the field.     

Comparison of Composition,Protein, Pasting,and Phenolic Compounds of Brown Rice and Germinated Brown Rice from Different Cultivars

Physical characteristics, amino acids composition, protein profiling, pasting characteristics, and phenolic compounds of brown rice (BR) and germinated brown rice (GBR) from different paddy cultivars (PB1, PS44, PB1509, PB1121, and PS5) were investigated. L* (lightness) decreased, but a* (redness and greenness) and b* (yellowness and blueness) increased with germination. Protein and ash content increased, whereas fat and amylose contents decreased with germination. GBR showed lower hardness and gumminess than BR. Foam stability and water absorption capacity from GBR flour were higher compared with BR flour. Accumulation of γ‐aminobutyric acid, histidine, arginine, proline, methionine, and acidic amino acids increased significantly with germination, and increase was related to change in accumulation of glutelin and prolamins. The accumulation of prolamins and glutelin acidic and basic subunits decreased with germination. GBR flour showed lower pasting viscosities compared with BR flour. Ferulic acid, p‐coumaric acid, and quercetin were present in both fractions of the bound form. GBR showed improved nutritional quality that varied in different cultivars. PB1121 was observed to be the best for producing GBR owing to greater changes brought in protein content, essential amino acids, catechin, chlorogenic acid, protocatechuic acid, vanillic acid, and foam stability.     

Indicated detection of two unapproved transgenic rice lines contaminating vermicelli products

We analyzed the DNA fragments extracted from four rice vermicelli products. The Bacillus thuringiensis (Bt) rice line, which has a construct similar to the GM Shanyou 63 line, was detected in some vermicelli products by identification of the junction region sequence between rice Act1 promoter and the Cry1Ac gene, and that between Cry1Ac and nos. In addition, we also detected a different Bt rice line by means of the junction region sequence between the maize ubiquitin promoter and cry1Ab gene and that between the cauliflower mosaic virus 35S promoter and the hygromycin phosphotransferase in some vermicelli products. Accordingly, we for the first time have detected the two transgenic Bt rice lines contaminating rice vermicelli samples. Furthermore, we developed a duplex real-time polymerase chain reaction (PCR) method for the simultaneous detection of both Bt rice lines.     

Kinetics and mechanism of free fatty acid formation on the surface of milled rice

The presence of free fatty acid (FFA) is an important factor in determining rice quality for brewing. FFA formation in milled rice during storage was monitored, and a two-parameter semiempirical kinetic model giving product concentration as a function of time is proposed to describe FFA formation on milled rice during storage. The model was tested using sets of data obtained from partially milled rice samples stored at 24, 37, and 50 degrees C and fully milled rice stored at 37 degrees C and 70% relative humidity. The predicted values provide very good fits (R(2) >or= 97%) of the experimental data at all storage temperatures. A two-substrate reaction mechanism representing a two-phase process is also presented. Milled rice FFA at a given storage time varied with storage temperatures. The kinetic model and mechanisms proposed could be useful in describing and predicting FFA contents of milled rice during storage and transportation.     

Quality and Agronomic Effects of Three High‐Molecular‐Weight Glutenin Subunit Transgenic Events in Winter Wheat

Quality and agronomic effects of three transgenic high molecular weight glutenin subunit (HMW‐GS) events were characterized in advanced‐generation breeding lines of hard winter wheat (Triticum aestivum L.) in three Nebraska crop years. Two of the transgenic events studied, Dy10‐E and B52a‐6, overexpress HMW‐GS 1Dy10, while the third event, Dx5 +Dy10‐H, overexpresses HMW‐GS 1Dx5 and, to a much lesser extent, 1Dy10. In addition, novel proteins possessing solubility characteristics defined as HMW‐GS were present in Dx5+Dy10‐H and B52a‐6. Average grain yield of lines derived from the three transgenic events was statistically lower than that of a group of control cultivars and advanced breeding lines, but not lower than the mean values of respective nontransgenic siblings. Grain hardness was influenced by one of the events. Dx5+Dy10‐H produced harder kernels than controls, its nontransgenic siblings, and the two additional transgenic events. All three events produced doughs with unusual mixing properties, although not likely to be directly useful in commercial applications. As a consequence, loaf volumes were depressed to variable degrees by the three events. The results indicated that over‐expression of HMW‐GS could eventually lead to improved breadmaking quality by optimizing the level of overexpression or by development and characterization of additional events.     

新型cry7Ab基因的鉴定克隆、表达与杀虫活性

本文应用PCR-RFLP法鉴定出Bt菌株HQ40中含有cry7Ab基因,并根据cry7A全长基因序列设计特异性引物,成功克隆了该基因。该基因核苷酸序列已经在国际基因库GeneBank中登记,其登录号为EU380678,并由Bt δ-内毒素基因国际命名委员会正式命名为cry7Ab4。通过穿梭载体pSTK 将该基因导入Bt 无晶体突变株中,获得工程菌HD7Ab4。SDS-PAGE分析表明cry7Ab4 基因在其中能正常表达,并形成菱形晶体。提取工程菌HD7Ab4和野生菌HQ40晶体蛋白,并在体外用胰蛋白酶酶解活化。分别对直翅目、鳞翅目和鞘翅目的害虫进行了杀虫活性测定。生测结果表明：Cry7Ab4蛋白trypsin酶解液对鞘翅目的大猿叶甲显示了一定的杀虫活性,其野生菌蛋白及表达蛋白酶解液LC50分别为231.59µg/ml及293.79µg/ml。表达产物虽不能使鳞翅目的小菜蛾、甜菜夜蛾和亚洲玉米螟死亡,但对它们的生长发育有明显的体重抑制作用。另外对马铃薯甲虫以及榆蓝叶甲也有体重抑制作用,而对直翅目的东亚飞蝗无毒。     

cry1Ac/cpti转基因大米水稻土中微生物群落的多样性研究

Two types of cry1Ac/cpti transgenic rice(GM1 and GM2)and their parental non-cry1Ac/cpti rice(CK1 and CK2)were planted in the field at Wufeng,Fujian Province,China for four years to investigate the influence of genetically modified rice on diversity of bacterial and fungal community in the paddy soil.The community composition and abundance of bacteria or fungi in the paddy soil were assessed at different growth stages of rice by denaturing gradient gel electrophoresis and real-time polymerase chain reaction based on 16S rRNA gene or SSU rRNA gene in the 4th year after the experimental establishment.The composition of bacterial or fungal community changed during rice growth,while no significant differences were observed between the fields cultivated with GM1and CK1,or between the fields cultivated with GM2 and CK2 in either bacterial or fungal community composition.The copy numbers of bacterial 16S rRNA gene in the soils with CK1,CK2,GM1 and GM2 ranged from 5.64×1011to 6.89×1011copies g-1dry soil at rice growth stages,and those of fungal SSU rRNA gene from 5.24×108to 8.68×108copies g-1dry soil.There were no marked differences in the copies of bacterial 16S rRNA gene or fungal SSU rRNA gene between CK1 and GM1 or between CK2 and GM2at any growth stage of rice.Planting cry1Ac/cpti transgenic rice had no significant effect on composition and abundance of bacterial and fungal community in paddy soil during the rice growing season at least in the short term.     

泥鳅生长及抗氧化 解毒酶系统对水体中转Cry1Ab/Ac基因水稻残遗物的响应

在泥鳅养殖水体中添加稻秆粉模拟水稻残遗物生境,研究了泥鳅生长和肝胰脏抗氧化酶(SOD、CAT)与解毒酶(GST)活性对转Cry1Ab/Ac基因水稻‘华恢1号’(HH1)的响应。设计以HH1稻秆粉10 mg·L?1、50 mg·L?1、100 mg·L?1和200 mg·L?1 4个梯度浓度处理泥鳅为试验组,以非转Bt基因水稻‘明恢63’(MH63)稻秆粉处理组为阴性对照,不加稻秆粉的基础饲养组为空白对照。结果显示:在4种稻秆粉浓度下,HH1组与MH63对照组泥鳅的特定生长率、肥满度、内脏系数及SOD、CAT和GST酶活性均无显著差异(P0.05);与空白对照比较,稻秆粉浓度升高对泥鳅生长的抑制逐渐增强,当浓度达到200 mg·L?1时,HH1组和MH63对照组泥鳅的特定生长率、内脏系数与CAT活性降低。研究结果表明,水体中低含量的转融合基因Cry1Ab/Ac水稻HH1稻秆粉对泥鳅的生长与生理酶活性没有明显影响,高浓度HH1和MH63稻秆粉均使泥鳅的生长和生理酶活性显著降低,这可能与养殖水体中浓度较高的悬浮稻秆粉妨碍了泥鳅的呼吸和滤食,及稻秆粉的分解降低了水体p H和溶氧量有关。