转基因抗除草剂油菜对十字花科杂草的基因漂移

以转基因抗除草剂油菜Q3为花粉供体材料,油菜远缘杂草为花粉受体材料,在自然传粉和人工辅助授粉条件下研究甘蓝型油菜与十字花科杂草间的基因漂移频率。结果表明,以转基因油菜为父本,十字花科杂草荠菜、碎米荠、播娘蒿、诸葛菜、风花菜、遏蓝菜和菜为母本,杂交高度不亲和,基因漂移率为0 % ,无生态风险,但对野芥菜的基因漂移率高达0 .885 %。野芥菜是我国大部分地区的常见杂草,种类繁多,分布范围广,大面积种植转基因抗除草剂油菜对野生芥菜的基因污染应引起高度重视。     

转基因抗除草剂油菜对近缘作物的基因漂移

以转基因抗除草剂油菜 Q3和 HCN- 19为花粉供体材料 ,油菜近缘作物为花粉受体材料 ,在自然授粉条件下研究甘蓝型油菜与芸薹属近缘作物间的基因漂移频率。结果表明 ,油菜对芸薹属 6个种甘蓝、黑芥、埃芥、芥菜型油菜、白菜型油菜和甘蓝型油菜的基因漂移率分别为 0、0 .0 2 4 %～ 0 .2 4 3%、 0 .0 2 8%～ 0 .0 92 %、 0 .10 9%～ 0 .95 1%、 0 .4 79%～ 0 .879%、 1.2 5 2 %～2 .191%。且基因漂移频率受多种因素影响 ,其中与杂交亲和性、花期同步率、种植面积等高度相关。通过花粉将抗除草剂基因漂移给近缘作物 ,油菜是需要特别关注的作物     

转基因水稻外源基因的漂移研究

转基因水稻基因漂移可能带来环境安全性问题。利用农杆菌介导,把hpt基因转化水稻品种,通过后代筛选,以稳定遗传的含单拷贝转化株系为转基因花粉供体材料,研究转基因水稻向非转基因水稻不育系和常规稻(花粉受体)的外源基因漂移频率。结果表明,相邻种植时转基因水稻向雄性不育系品种的漂移频率为31.74%。随距离的增加漂移频率明显下降,在26m处仍能够检测到含外源基因个体的存在,并且在距离4m处出现一个漂移频率的升高;转基因水稻向常规品种的漂移频率则明显低于不育系,一般在2.0%以下,随距离的增加也呈现下降的趋势,在18m处开始无法检测到含外源基因的个体。     

转基因玉米外源基因通过花粉漂移的频率和距离

基因漂移是转基因生物可能引起生态环境安全性问题的主要风险之一,选用美国孟山都公司的转基因抗除草剂玉米GA21进行了外源基因向周边环境遗传漂移的距离和频率的研究,结果表明:转基因玉米的外源基因可以向周边玉米品种进行漂移,其最大漂移频率为45.10%,150m处仍能检测到外源基因的漂移,防止转基因玉米基因向外扩散的最佳途径为设置隔离带,隔离距离以200m以上为好。     

水稻移栽密度和早期水管理对无芒稗生态控制效果的影响

在 5 0株·m-2 无芒稗的干扰下 ,杂交水稻Ⅱ优 2 0 70移栽密度 1 9 8cm× 2 6 4cm处理对无芒稗的生态控制效果显著高于较稀密度的 1 9 8cm× 4 6 2cm ,无芒稗颖果减少 71 %。移栽密度 1 9 8cm× 2 6 4cm保证了较高的水稻有效穗和千粒重 ,比移栽密度 1 9 8cm× 4 6 2cm的水稻增产 1 3 3%。早期深水灌溉能有效控制萌芽期的无芒稗 ,但不能防治 3叶龄的无芒稗     

转基因白桦的花粉活力及外源基因的遗传表达分析

采用荧光素二乙酸酯（FDA）染色比较分析转基因白桦花粉的活力的结果表明,转基因白桦的花粉活力均低于非转基因白桦。应用多重PCR和Northern杂交技术分析表明,转基因白桦花粉中有外源基因整合和转录表达。检测转基因白桦花粉中gus报告基因的活性,显示gus报告基因在花粉表达。     

转基因插入对水稻花粉活力和杂交结实的影响

以花粉活力和杂交结实率评价转基因插入对水稻品种异交潜力的影响.选用含bar、crylAb、BADH和Xa21等4种基因的5份转基因水稻品系研究转基因插入对转基因受体品种花粉离体萌发率的影响.结果表明,不同受体品种花粉离体萌发率差异显著;转基因水稻花粉离体萌发率在0.416～0.584间,与受体品种(0.004～0.574)相近;转基因品种与相应受体品种间花粉离体萌发率差异不显著.对所选配的26个人工杂交组合杂种结实调查表明,bar、crylAb基因的插入对受体品种杂交结实率影响显著,而Xa21基因的插入对受体品种杂交结实率影响较小;转基因水稻品种(花粉供体)与非转基因水稻品种的杂交结实率变幅为0.056～0.13,在受体品种(花粉供体)与非转基因水稻品种的杂交结实率范围(0.052～0.417)之内.本研究花粉离体萌发和杂交结实结果表明转基因的插入对水稻品种异交潜力的影响甚微.     

花粉离体培养基的优化和苯达松除草剂对敏感型水稻恢复系'Mc526'花粉活力的影响

建立了适合水稻花粉离体萌发的液体培养基配方,并用花粉萌发和碘-碘化钾（I2-KI）染色测定法检测了喷施苯达松后含敏感致死基因的水稻恢复系‘Mc526’的花粉生活力。结果表明,苯达松处理前期（施后14d）,‘Mc526’花粉活力与未作处理的接近;处理后期（施后5-8d）略低于未作处理的,但差异不显著。苯达松对母本结实率影响不显著。     

UV-B辐射增强对抗除草剂转基因水稻 CH4排放的影响

在大田条件下,研究了UV-B(ultraviolet-B)辐射增强对抗除草剂转基因水稻及亲本常规水稻甲烷(CH4)排放的影响。UV-B辐射设2水平,即对照(CK,自然光),增强(Elevated,14.4 kJ·m-·2d-1),相当于南京地区大气臭氧耗损25%的辐射剂量。结果表明,UV-B辐射增强并没有改变稻田CH4排放通量的季节性变化规律。与对照相比,UV-B辐射增强显著提高CH4排放通量和累积排放量。水稻分蘖期CH4累积排放量最高,占全生育累积排放量的51.55%—61.01%;其次是拔节至孕穗期,占20.00%—26.64%。抗除草剂转基因水稻的CH4排放通量和累积排放量显著低于亲本常规水稻。研究说明,UV-B辐射增强下种植抗除草剂转基因水稻对于减缓稻田甲烷排放有积极意义。     

抗除草剂转基因水稻对二种刺吸性害虫和二种天敌的影响

为探明转基因抗除草剂水稻对非目标生物的影响,于2007年和2008年对转Bar基因抗除草剂籼稻Bar68-1和非转基因对照D68稻田的2种常见刺吸性害虫和2种天敌在水稻分蘖中期、分蘖末期、齐穗期和乳熟期4个时期的种群密度进行了调查和比较。结果显示,抗除草剂水稻与对照稻田的飞虱、叶蝉、盲蝽、蜘蛛的发生密度,除不喷药处理区的2007年水稻乳熟期和2008年分蘖中期的盲蝽科的发生密度差异显著外,其余均无显著差异。试验结果表明,转基因抗除草剂水稻Bar68-1对包括飞虱、叶蝉、盲蝽、蜘蛛等类群在内的稻田非目标生物的田间自然种群的影响与非转基因水稻基本相同。     

两种温度下模拟氮沉降对陆稻与稗草竞争的影响

在昼/夜温度为35 ℃/25 ℃和30 ℃/20 ℃条件下,研究了模拟氮沉降对稗草和陆稻生长及竞争关系的影响.结果表明：35 ℃/25 ℃条件下,每年输入4 g·m^-2氮处理,稗草和陆稻地上部分生物量分别比对照增加29.18%和27.80％,稗草吸收氮和磷量分别增加87.33%和49.73%,而陆稻吸收氮和磷量无显著变化.在30 ℃/20 ℃条件下,每年输入2、4、6 g·m^-2氮处理后,稗草地上部分生物量分别比对照增加48.99％、72.68％和36.18％,分蘖数分别增加111.11％、122.22％和144.44％,稗草植株氮和磷吸收量分别增加108.88％、129.22%、134.29％和16.53％、65.05%、22.47%,而陆稻均无显著变化.在30 ℃/20 ℃条件下,氮沉降显著增加了稗草与陆稻地上部分生物量的比值,但在35 ℃/25 ℃条件下影响不显著.表明氮沉降增加可能会提高稗草而降低陆稻的竞争力,而且在温度较低的情况下,这种趋势更明显.     

A large-scale field study of transgene flow from cultivated rice (Oryza sativa) to common wild rice (O. rufipogon) and barnyard grass (Echinochloa crusgalli)

The introgression of transgenes into wild relatives or weeds through pollen-mediated gene flow is a major concern in environmental risk assessment of transgenic crops. A large-scale (1.3–1.8 ha) rice gene flow study was conducted using transgenic rice containing the bar gene as a pollen donor and Oryza rufipogon as a recipient. There was a high frequency of transgene flow (11%−18%) at 0–1 m, with a steep decline with increasing distance to a detection limit of 0.01% by 250 m. To our knowledge, this is the highest frequency and longest distance of gene flow from transgenic rice to O. rufipogon reported so far. On the basis of these data, an adequate isolation distance from both conventional and transgenic rice should be taken for in situ conservation of common wild rice. Meanwhile, there is no evidence of transgene introgression into barnyard grass, even when it has coexisted with transgenic rice containing the bar gene for five successive years. Thus, the environmental risk of gene flow to this weedy species is of little concern.     

转基因水稻基因流的发生与生态学后果

中国是水稻的起源中心之一,分布着丰富的野生种质资源.自转基因水稻获得安全证书以来,转基因水稻与其近缘野生种间的基因流受到广泛关注.本文对转基因水稻基因流的发生及其可能引起的生态学后果进行了综述和展望.认为转基因水稻能够与栽培稻、野生稻、杂草稻、稗草等成功杂交,但基因流发生频率较低且变化较大.基因流成功发生后,由于转基因水稻具转基因新性状而有适合度优势,转基因可能只通过少数几代就进入野生种群.当转基因植株进入野生种群并在自然条件下长期存在时,转基因植株与近缘野生种间的竞争关系和相对适合度将决定混合种群的动态变化.研究转基因水稻基因流的影响及其长期生态学后果对合理保护与利用野生种质资源具有重要意义.     

转基因水稻外源基因向近缘种群漂流和渐渗的研究进展

水稻是我国最重要的粮食作物之一,我国有8亿以上的人口以稻米作为主食。但在水稻生产中,由于病、虫、草害及不良气候等逆境因子的影响,严重制约了水稻的高产、稳产。转基因生物技术的迅速发展,为水稻抗性育种提供了新途径。自20世纪80年代以来,我国全方位地开展了转基因水稻的研发,目前已经培育出大量的抗病、抗虫、抗除草剂和抗逆的转基因水稻品种,这将为提高我国水稻的生产力和确保粮食安全做出重要的贡献。但转基因水稻的基因漂流及其可能带来的生物安全问题备受关注。已有报道证明,外源转基因可以通过异交向非转基因品种和野生近缘种漂流。在不同的试验条件下,抗除草剂基因有0．05％-0．53％逃逸的可能,其向不育系的最大漂移频率可达4．518％。抗虫基因向相邻非转基因水稻的平均漂移频率最高为0．875％。因此,本文对水稻与其近缘野生种的杂交情况,转基因水稻外源基因向非转基因品种、野生近缘种以及野生非近缘种的漂流和渐渗及其潜在的生态环境风险等方面进行了简要分析,并对转基因水稻的发展进行了展望,以期为转基因水稻的安全应用提供参考。     

花粉介导的转Bt基因棉花田间基因流监测

采用花粉粒染色法对转Bt基因棉的花粉漂移距离和强度进行了观测,并应用PCR法检测转Bt基因棉的基因流频率.花粉粒染色法监测结果表明：同株异花间的花粉散布频率显著高于异株异花间(P<0.01);靠近转Bt基因棉花粉染色区1 m处的平均花粉散布频率,在东、南、西、北4个方向分别为44.8%、48.9%、57.1%和21.5%,但随着距转基因棉田距离的增大,4个方向的平均花粉散布频率都呈下降趋势.PCR结果的统计分析表明,在25m内,花粉散布距离和方向对基因流频率有极显著影响(P<0.01),随着距转基因棉田距离的增大,基因流频率呈下降趋势,最远距离为25 m时的最高基因流频率为2.0%.     

Dramatic reduction of crop-to-crop gene flow within a short distance from transgenic rice fields

Genetically modified (GM) rice with enhanced agronomic traits and pharmaceutical uses are ready for widespread adoption. Little is known about isolation requirements for achieving stringent transgene confinement in rice. To investigate the extent of pollen-mediated crop-to-crop transgene flow, we conducted a field experiment with four plot-size treatments of adjacent GM and nonGM rice (Oryza sativa) in China. Three insect-resistant GM rice (Bt/CpTI) and nonGM isogenic lines were used in the study. The hygromycin-resistance transgene (hpt) marker was used to screen seeds from the nonGM rice rows at different distance intervals from GM rice plots. Based on the examination of > 2.1 million germinated seeds, we found a dramatic reduction in transgene frequencies with increasing distance from the GM crop, ranging from c. 0.28% at 0.2 m to < 0.01% at 6.2 m. In addition, different plot size did not significantly affect the frequencies of gene flow. In conclusion, pollen-mediated crop-to-crop transgene flow in rice can be maintained at negligible levels with short spatial isolation. The model can also be applied to other crops with self- and wind-pollination.     

Effect of boron on the pollen vitality and yield of rice plants (Oryza sativa L. var. Jaya)

Summary Rice plants were grown in sand cultures with nutrient solution including boron at 0, 1, 2.5 and 5 ppm concentrations.In general, supply of boron improved the pollen vitality of rice flowers. It was stimulating up to 2.5 ppm concentration in the nutrient solution, beyond which (i.e., at 5 ppm) inhibitory effects appeared. The availability of boron also increased the yield of rice grains in the same order.Stimulating effects of boron may be linked with greater availability of sugars, increased enzymatic activity and respiration which favoured better growth of pollen. Inhibitory effects of stronger concentration of boron (5 ppm) may be related with physiological depression and injury to protoplasm itself.     

氮素增加条件下丛枝菌根真菌对陆稻和无芒稗相互作用的调节

采用盆栽方法研究了增氮条件下丛枝菌根真菌(AMF)对无芒稗(Echinochloa crusgallivar.mitis L.)和陆稻(Oryza sativa L.)相互作用的调节.结果表明:在单种条件下,无芒稗的菌根侵染率增加,陆稻的菌根侵染率降低;无AMF和接种AMF处理陆稻的生物量分别提高13.48%和42.35%,总磷吸收分别提高2.55%和4.07%,总氮吸收分别提高62.09%和30.35%;无芒稗的生物量分别提高15.65%和20.24%,总磷吸收分别提高4.06%和3.88%,总氮吸收分别提高30.35%和15.10%.在混种条件下,无芒稗的菌根侵染率增加,而陆稻无显著变化;无AMF和接种AMF的无芒稗与陆稻的生物量比值降低,总氮吸收比值无显著变化,无AMF处理的总磷吸收比值增加,而接种AMF处理降低.表明增氮条件下AMF提高了无芒稗对陆稻的竞争.     

Transcriptional silencing and developmental reactivation of cry1Ab gene in transgenic rice

One transgenic rice line lacking CrylAb expression product was screened in the progenies of Agrobacterium-transformed transgenic rice variety Zhong 8215 with a cry1Ab gene under field releasing conditions by using GUS histochemical assay and Western blot. Molecular hybridization results revealed that the crylAb gene was silenced in the transgenic rice variety Zhong 8215 and two copies of ubiquitin promoter were integrated into the rice genome. The silencing of crylAb gene in transgenic rice was found to be due to the methylation of the ubiquitin promoter as revealed by methylation analysis. Meanwhile, different concentrations of demethylation reagent 5-azacytidine combining with different treatment time were employed to treat the silenced transgenic rice seeds. The results indicated that 5-azacytidine could reactivate 8%-30% of the silenced transgenic rice plants and the expression level of the reactivated cry1Ab transgene could reach as high as 0.147% of the total soluble protein. Treatment with low con     

Expression of betaine aldehyde dehydrogenase gene and salinity tolerance in rice transgenic plants

Betaine as one of osmolytes plays an important role in osmoregulation of most high plants. Betaine aldehyde dehydrogenase C BADH) is the second enzyme involved in betaine biosynthesis. The BADH gene from a halophite, Atriplex hortensis, was transformed into rice cultivars by bombarment method. Totally 192 transgenic rice plants were obtained and most of them had higher salt tolerance than controls. Among transgenic plants transplanted in the saline pool containing 0.5% NaCl in a greenhouse, 22 survived, 13 of which set seeds, and the frequency of seed setting was very low, only 10% . But the controls could not grow under the same condition. The results of BADH ac-tivity assay and Northern blot showed that the BADH gene was integrated into chromosomes of transgenic plants and expressed.