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     

A field study of pollen-mediated gene flow from Mediterranean GM rice to conventional rice and the red rice weed

The objective of this study was to assess the frequency of pollen-mediated gene flow from a transgenic rice line, harbouring the gusA and the bar genes encoding respectively, -glucuronidase and phosphinothricin acetyl transferase as markers, to the red rice weed and conventional rice in the Spanish japonica cultivar Senia. A circular field trial design was set up to investigate the influence of the wind on the frequency of pollination of red rice and conventional rice recipient plants with the transgenic pollen. Frequencies of gene flow based on detection of herbicide resistant, GUS positive seedlings among seed progenies of recipient plants averaged over all wind directions were 0.036 ± 0.006% and 0.086 ± 0.007 for red rice and conventional rice, respectively. However, for both red rice and conventional rice, a clear asymmetric distribution was observed with pollination frequency favoured in plants placed under the local prevailing winds. Southern analyses confirmed the hemizygous status and the origin of the transgenes in progenies of surviving, GUS positive plants. Gene flow detected in conventional rice planted at 1, 2, 5 and 10 m distance revealed a clear decrease with increasing distance which was less dramatic under the prevailing wind direction. Consequences of these findings for containment of gene flow from transgenic rice crops to the red rice weed are discussed. The precise determination of the local wind conditions at flowering time and pollination day time appear to be of primary importance for setting up suitable isolation distances.     

Segregation for endosperm lysine in F2, F3 and F4 progeny from a cross of in vitro-selected and unselected cultivar of rice

Summary Lysine is a limiting amino acid for optimal nutritional quality in rice grain. In vitro selections using inhibitory levels of lysine plus threonine or s-aminoethylcysteine allow the predictable recovery of variants with elevated levels of lysine and protein. These methods may generate useful starting germplasm for plant breeders. This study was conducted to define the genetics of lysine mutants in progeny from crosses of mutants derived from cells cultured in vitro in the presence of inhibitory levels of lysine plus threonine and s-(2-aminoethyl)-cysteine. In vitro selections produce a wide range of mutants, including endosperm mutants with elevated lysine and protein levels as well as mutants for high and low seed weights. Mutants were analyzed for lysine content by the endosperm half-seed method in which the halves without the embryo were ground and acid hydrolyzed for amino acid determinations. The halves with the embryos were preserved for later germination. In two different F₂ populations derived from a cross of a selected mutant x M-101, a parental marker, there was an inverse relationship between seed weight and percent lysine in endosperm protein (R² 0.52 and 0.56). The F₂ segregation patterns show that elevated lysine is inherited as a recessive gene and that increased lysine is correlated with decreased seed size. F₃ and F₄ data provide evidence for the transmission of high lysine genes to advanced germplasm in rice. This work supports our earlier conclusions that high lysine phenotypes can be recovered predictably from in vitro selections. The elevated lysine phenotypes are frequently, but not exclusively, associated with opaque seed. Some segregants from crosses produced increased lysine in plants with near normal seed weight and good fertility.Research done under the auspices of the USDA, ARS, Plant Sciences Institute, Plant Molecular Biology Laboratory, Beltsville, MD 20705, USA     

Effects of biotechnology on biodiversity: herbicide-tolerant and insect-resistant GM crops

Biodiversity is threatened by agriculture as a whole, and particularly also by traditional methods of agriculture. Knowledge-based agriculture, including GM crops, can reduce this threat in the future. The introduction of no-tillage practices, which are beneficial for soil fertility, has been encouraged by the rapid spread of herbicide-tolerant soybeans in the USA. The replacement of pesticides through Bt crops is advantageous for the non-target insect fauna in test-fields. The results of the British Farm Scale experiment are discussed. Biodiversity differences can mainly be referred to as differences in herbicide application management.     

Gene flow from transgenic rice to red rice (Oryza sativa L.) in the field

In this study, we simulate a transgenic rice crop highly infested with red rice to examine transgene transfer from a transgenic line (A2504) resistant to glufosinate ammonium to cohabitant red rice. The red rice was sown along with the transgenic line at the highest density found in naturally infested crops in the region. Agricultural practices similar to those used to control red rice infestation in northern Italy rice fields were used to reproduce the local rice production system. During the first 2 years, the field was treated with herbicide at the appropriate time; in the first year the dosage of herbicide was three times the recommended amount. In this first year, detectable red rice plants that escaped herbicide treatment were manually removed. Nevertheless, two herbicide‐resistant hybrid plants (named 101 and 104) were identified in the experimental field during the second year of cultivation. Phenotypic and molecular characterisation suggests the hybrid nature of these two plants, deriving from crossing events involving A2504, respectively, with red rice (plant 101) and the buffer cultivar Gladio (plant 104). The progeny of two subsequent generations of the two plants were examined and the presence of the transgene detected, indicating stable transfer of the transgene across generations. In conclusion, despite control methods, red rice progeny tolerant to the herbicide can be expected following use of transgenic rice and, consequently, difficulties in controlling this weed with chemicals will emerge in a relatively short time.     

落羽杉与墨西哥落羽杉3个杂交后代品系的生长和杂种优势分析

以落羽杉〔Taxodium distichum(Linn.)Rich.〕和墨西哥落羽杉(T.mucronatum Tenore)及其3个杂交后代品系'中山杉405'('Zhongshansha 405')、'中山杉406'('Zhongshansha 406')和'中山杉407'('Zhongshansha 407')2年生盆栽扦插苗为研究对象,对株高和地径的年增长量、总叶面积、株高和地径的增长量动态变化、株高和地径的生长期进行了比较分析,并根据地径增长量分析了3个杂交后代品系的杂种优势.结果表明:3个杂交后代品系的年株高增长量介于父本与母本之间但无显著差异,而三者的年地径增长量和总叶面积则明显高于父本和母本.墨西哥落羽杉与3个杂交后代品系的株高和地径生长期均一致,分别为5月11日至10月5日和6月1日至11月9日;而落羽杉的株高和地径生长期分别为5月11日至8月10日和6月8日至11月16日.5月份至10月份,墨西哥落羽杉的株高持续增长,而3个杂交后代品系的株高不连续增长,落羽杉的株高则在8月10日之后停止生长;生长初期和生长末期,3个杂交后代品系的地径增长量均高于父本和母本;秋季3个杂交后代品系均能快速进行地径生长,而此时父本和母本的地径增长量均下降.除夏季的少数时间段外,'中山杉405'在整个生长季都表现出明显的杂种优势,而'中山杉406'和'中山杉407'在整个生长季总体上表现出杂种优势,且它们的杂种优势最大值均出现在生长后期,说明供试的3个杂交后代品系在秋季也能快速生长.研究结果显示:供试的3个杂交后代品系均遗传了落羽杉地径生长迅速和墨西哥落羽杉株高生长期长的特性,秋季3个杂交后代品系的地径和株高增长量总体上均高于父本和母本,表现出明显的杂种优势.     

Molecular breeding of a novel herbicide-tolerant rice by gene targeting

We have previously reported the production of a rice cell line tolerant to the acetolactate synthase (ALS)-inhibiting herbicide bispyribac (BS), and demonstrated that the BS-tolerant phenotype was due to a double mutation in the rice ALS gene. We further indicated that while changing either of the two amino acids (W548 L or S627I) individually resulted in a BS-tolerant phenotype, conversion of both amino acids simultaneously conferred increased tolerance to BS. As the BS-tolerant cell line had lost the ability to regenerate during two years of tissue culture selection, we attempted to introduce these two point mutations into the rice ALS gene via gene targeting (GT). Using our highly efficient Agrobacterium-mediated transformation system in rice, we were able to regenerate 66 independent GT rice plants from 1500 calli. Furthermore, two-thirds of these plants harbored the two point mutations exclusively, without any insertion of foreign DNA such as border sequences of T-DNA. The GT plants obtained in the present study are therefore equivalent to non-GM herbicide-tolerant rice plants generated by conventional breeding approaches that depend on spontaneous mutations. Surprisingly, GT rice homozygous for the modified ALS locus showed hyper-tolerance to BS when compared to BS-tolerant plants produced by a conventional transgenic system; ALS enzymatic activity in plants homozygous for the mutated ALS gene was inhibited only by extremely high concentrations of BS. These results indicate that our GT method has successfully created novel herbicide-tolerant rice plants that are superior to those produced by conventional mutation breeding protocols or transgenic technology.     

Mapping quantitative trait loci in F2 incorporating phenotypes of F3 progeny

In plants and laboratory animals, QTL mapping is commonly performed using F(2) or BC individuals derived from the cross of two inbred lines. Typical QTL mapping statistics assume that each F(2) individual is genotyped for the markers and phenotyped for the trait. For plant traits with low heritability, it has been suggested to use the average phenotypic values of F(3) progeny derived from selfing F(2) plants in place of the F(2) phenotype itself. All F(3) progeny derived from the same F(2) plant belong to the same F(2:3) family, denoted by F(2:3). If the size of each F(2:3) family (the number of F(3) progeny) is sufficiently large, the average value of the family will represent the genotypic value of the F(2) plant, and thus the power of QTL mapping may be significantly increased. The strategy of using F(2) marker genotypes and F(3) average phenotypes for QTL mapping in plants is quite similar to the daughter design of QTL mapping in dairy cattle. We study the fundamental principle of the plant version of the daughter design and develop a new statistical method to map QTL under this F(2:3) strategy. We also propose to combine both the F(2) phenotypes and the F(2:3) average phenotypes to further increase the power of QTL mapping. The statistical method developed in this study differs from published ones in that the new method fully takes advantage of the mixture distribution for F(2:3) families of heterozygous F(2) plants. Incorporation of this new information has significantly increased the statistical power of QTL detection relative to the classical F(2) design, even if only a single F(3) progeny is collected from each F(2:3) family. The mixture model is developed on the basis of a single-QTL model and implemented via the EM algorithm. Substantial computer simulation was conducted to demonstrate the improved efficiency of the mixture model. Extension of the mixture model to multiple QTL analysis is developed using a Bayesian approach. The computer program performing the Bayesian analysis of the simulated data is available to users for real data analysis.     

培矮64S/爪哇稻F1农艺性状表现和对照优势分析

本文报道了利用原始爪哇稻资源与光温敏雄性不育系培矮64S配制的27个籼爪交组合在长沙的农艺性状和杂种优势表现.从总体上来看,籼爪交组合与对照相比在每穗实粒数和理论产量上无显著差异,在其它性状上均有极显著的差异;籼爪交组合在秆高、穗长、每穗总粒数、每穗实粒数和千粒重方面有正向对照优势,在播始历期、有效分蘖数、结实率、理论产量和实际产量上存在负向对照优势.从个体上来看,籼爪交组合理论产量对照优势＞40%的比例为11.1%.实际产量对照优势＞40%的机率为3.7%,说明爪哇稻资源在籼爪交杂种优势利用中具有利用价值.本文还对爪哇稻资源在籼爪杂种优势利用中的一些问题进行了讨论.     

Epistatic gene effects on the yield of the parents of F1, F2, BC1 and BC2 progeny

The genetic analysis of 5 tomato hybrids (Danubius F₁, Luna F₁, Lido F₁, Balkan F₁ and Mi-10 F₁) was made. We produced their F₁, F₂, BC₁ and BC₂ generations and analysed their yield (on the first three flower branches) as well as some of the yield components of tomato fruits (mean fruit weight, mean fruit weight on the first flower branch, fruit length, fruit width, and number of locules). In order to estimate the gene effects, we applied the additive-dominance mode with three and six parameters. Epistatic gene effects were estimated by applying the six-parameter mode (Mather and Jinks 1982). As for yield and yield components, there were significant differences between the mean values of parents and their progeny. On the basis of the investigated genetic parameters, the obtained results suggested that the additive and dominance gene effects prevailed in the yield and yield components (Danubius F₁, Luna F₁, Lido F₁, Mi-10 F₁), whereas epistatic gene effects were excluded. As for the hybrid Balkan F₁, we recorded significant gene effects, both the additive and the dominance ones in the yield inheritance: additive x additive and dominance x dominance (with the negative sign). The estimated values of the epistatic gene effects were the most prominent in inheriting the feature average fruit weight on the first flower branch — additive x dominance gene effects. They represented the most frequent type of the interallele interaction recorded in the investigated hybrids.     

Cytogenetic studies on two F1 hybrids of autotetraploid rice varieties showing extremely high level of heterosis

Mechanisms of two F1 hybrids (D46A × DTP-4 and D46A × Dminghui63) of autotetraploid rice (2n = 4x = 48) showing extremely high pollen fertility 87.40% and 85.97%, respectively, seed set 82.00% and 79.00%, respectively and extremely high level of heterosis were analyzed cytologically. The chromosome pairing of D46A × DTP-4 and D46A × Dminghui63 was normal at metaphase I(MI), and had almost no I or III, with an average of 0.020I +14.36 II 6.44rod+7.91ring) +0.01III + 4.80 IV + 0.01VIII and 0.06 I + 17.67 II (11.01rod + 6.67ring)] + 0.06 III +3.10IV+0.01VI, respectively. The most frequent chromosome configurations were 10II+7IV and 12II+bIV. The bivalent frequency was less frequent in hybrids than that in restoring parents, and the same results were gained from univalents, trivalent and multivalents. However, the quadrivalent frequency was significantly higher in hybrids than that in restoring parents at MI. The other meiotic phases progressed normally, except for low percentages of PMCs with lagging chromosomes at AI and low percentages of PMCs with micronuclei at telophaseI (TI) and telophaseII (TII). PMCs with lagging chromosomes at AI and PMCs with micronuclei at TI and TII showed negative correlation between pollen fertility and seed set. Above 90% of the PMCs could form normal microspores, which resulted in the production of viable pollen grains, abnormal microspores were observed including penta-fission and hexa-fission. Based on these results we suggest that the two F1 hybrids had better behaviors of chromosome pairing and genetic stability than autotetraploid rice and other autotetraploid plants ever studied.     

The genetics ofPhytophthora infestans: Segregation of allozyme markers in F2 and backcross progeny and the inheritance of virulence against potato resistance genesR2 andR4 in F1 progeny

The purpose of this work was to investigate the inheritance of allozymes and virulence inPhytophthora infestans. For two previously known allozyme systems (glucose phosphate isomerase and peptidase) we discovered new alleles,Gpil (98) andPepl (83 and96). We also determined that a previously uncharacterized enzyme system, mannose phosphate isomerase, had at least two loci and four electromorphs (93, 100, 105, and 111) whose allelic relationships are not yet elucidated. We observed normal segregations in F1 progeny, but there were some unexpected segregations in backcrosses and sibcrosses. One backcross produced predominantly heterozygous progeny for bothGpil andPepl, with the expected classes of homozygotes virtually absent. An excess of heterozygotes forGpil was observed in another backcross, and in sibcrosses there were deficiencies of expected homozygous classes. In F1 progeny, virulence/ avirulence against potato resistance geneR2 segregated in a 1:1 ratio (P = 0.3), while the segregation of virulence/avirulence toR4 departed somewhat from a 1:1 ratio (P = 0.07). These results support the gene-for-gene hypothesis inP. infestans and suggest that virulence againstR2 is controlled at a single locus, while virulence againstR4 may be determined at one or two loci.     

Gene flow from cultivated rice (Oryza sativa) to its weedy and wild relatives

BACKGROUND AND AIMS: Transgene escape through gene flow from genetically modified (GM) crops to their wild relative species may potentially cause environmental biosafety problems. The aim of this study was to assess the extent of gene flow between cultivated rice and two of its close relatives under field conditions. METHODS: Experiments were conducted at two sites in Korea and China to determine gene flow from cultivated rice (Oryza sativa L.) to weedy rice (O. sativa f. spontanea) and common wild rice (O. rufipogon Griff.), respectively, under special field conditions mimicking the natural occurrence of the wild relatives in Asia. Herbicide resistance (bar) and SSR molecular finger printing were used as markers to accurately determine gene flow frequencies from cultivated rice varieties to their wild relatives. KEY RESULTS: Gene flow frequency from cultivated rice was detected as between approx. 0.011 and 0.046 % to weedy rice and between approx. 1.21 and 2.19 % to wild rice under the field conditions. CONCLUSIONS: Gene flow occurs with a noticeable frequency from cultivated rice to its weedy and wild relatives, and this might cause potential ecological consequences. It is recommended that isolation zones should be established with sufficient distances between GM rice varieties and wild rice populations to avoid potential outcrosses. Also, GM rice should not be released when it has inserted genes that can significantly enhance the ecological fitness of weedy rice in regions where weedy rice is already abundant and causing great problems.     

The effect of parental divergence on F2 heterosis in winter wheat crosses

Summary In winter wheat (Triticum aestivum L.), the development of a methodology to estimate genetic divergence between parental lines, when combined with knowledge of parental performance, could be beneficial in the prediction of bulk progeny performance. The objective of this study was to relate F₂ heterosis for grain yield and its components in 116 crosses to two independent estimates of genetic divergence among 28 parental genotypes of diverse origins. Genetic divergence between parents was estimated from (a) pedigree relationships (coefficients of kinship) determined without experimentation, and (b) quantitative traits measured in two years of field experimentation in Kansas and North Carolina, USA. These distances, designated (1 -r) and G, respectively, provided ample differentiation among the parents. The 116 F₂ bulks were evaluated at four locations in Kansas and North Carolina in one year. Significant rank correlations of 0.46 (P = 0.01) and 0.44 (P = 0.01) were observed between G and grain yield and kernel number heterosis, respectively. Although (1 -r) was poorly associated with grain yield heterosis, G and midparent performance combined to account for 50% of the variation in F₂ yields among crosses when (1 -r) was above the median value, whereas they accounted for only 9% of the variation among crosses when (1-r) was below the median. Midparent and (1 -r) had equal effects on F₂ grain yield (R ²= 0.40) when G was greater than the median value. A breeding strategy is proposed whereby parents are first selected on the basis of performance per se and, subsequently, crosses are made between genetically divergent parents that have both large quantitative (G) and pedigree divergence (1 -r).Paper No. 12162 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, NC 27695-7643, and Contribution No. 89-396-J of the Kansas Agricultural Experiment Station, Manhattan, KS 66506     

Understanding the genetic and molecular constitutions of heterosis for developing hybrid rice

The wide adoption of hybrid rice has greatly increased rice yield in the last several decades. The utilization of heterosis facilitated by male sterility has been a common strategy for hybrid rice development. Here, we summarize our efforts in the genetic and molecular understanding of heterosis and male sterility together with the related progress from other research groups. Analyses of F1 diallel crosses show that strong heterosis widely exists in hybrids of diverse germplasms, and inter-subsp...     

Importance of over-dominance as the genetic basis of heterosis in rice

In populations derived from commercial hybrid rice combination Shanyou 10, F1 hetero-sis and F2 inbreeding depression were observed on grain yield (GYD) and number of panicles (NP). Using marker loci evenly distributed on the linkage map as fixing factors, the F2 population was divided into sub-populations. In a large number of sub-populations, significant correlations were observed between heterozygosity and GYD, and between heterozygosity and NP. This was especially true in type III sub-populations in which the genotype of a fixing factor was heterozy-gotes. In type III sub-populations, 15 QTL for GYD and 13 QTL for NP were detected, of which the majority exhibited over-dominance effects for increasing the trait values. This study showed that over-dominance played an important role in the genetic control of heterosis in rice.