Xiaoling Song

Potential gene flow of two herbicide-tolerant transgenes from oilseed rape to wild B. juncea var. gracilis

Four successive reciprocal backcrosses between F1 (obtained from wild Brassica juncea as maternal plants and transgenic glyphosate- or glufosinate-tolerant oilseed rape, B. napus, as paternal plants) or subsequent herbicide-tolerant backcross progenies and wild B. juncea were achieved by hand pollination to assess potential transgene flow. The third and forth reciprocal backcrosses produced a number of seeds per silique similar to that of self-pollinated wild B. juncea, except in plants with glufosinate-tolerant backcross progeny used as maternal plants and wild B. juncea as paternal plants, which produced fewer seeds per silique than did self-pollinated wild B. juncea. Germination percentages of reciprocal backcross progenies were high and equivalent to those of wild B. juncea. The herbicide-tolerant first reciprocal backcross progenies produced fewer siliques per plant than did wild B. juncea, but the herbicide-tolerant second or third reciprocal backcross progenies did not differ from the wild B. juncea in siliques per plant. The herbicide-tolerant second and third reciprocal backcross progenies produced an amount of seeds per silique similar to that of wild B. juncea except for with the glufosinate-tolerant first and second backcross progeny used as maternal plants and wild B. juncea as paternal plants. In the presence of herbicide selection pressure, inheritance of the glyphosate-tolerant transgene was stable across the second and third backcross generation, whereas the glufosinate-tolerant transgene was maintained, despite a lack of stabilized introgression. The occurrence of fertile, transgenic weed-like plants after only three crosses (F1, first backcross, second backcross) suggests a potential rapid spread of transgenes from oilseed rape into its wild relative wild B. juncea. Transgene flow from glyphosate-tolerant oilseed rape might be easier than that from glufosinate-tolerant oilseed rape to wild B. juncea. The original insertion site of the transgene could affect introgression.

Agronomic performance of F1, F2 and F3 hybrids between weedy rice and transgenic glufosinate-resistant rice.

BACKGROUND Studies of hybrid fitness, of which agronomic performance may be an indicator, can help in evaluating the potential for introgression of a transgene from a transgenic crop to wild relatives. The objective of this study was to assess the agronomic performance of reciprocal hybrids between two transgenic glufosinate-resistant rice lines, Y0003 and 99-t, and two weedy rice accessions, WR1 and WR2, in the greenhouse. RESULTS F1 hybrids displayed heterosis in height, flag leaf area and number of spikelets per panicle. The agronomic performance of F1 between WR1 and Y0003 was not affected by crossing direction. The tiller and panicle numbers of F1 individuals were higher than their F2 counterparts. However, these traits did not change significantly from the F2 to the F3 generation or in hybrids with weedy rice as maternal or paternal plants. For all hybrids, the in vitro germination rates of fresh pollen were similar and significantly lower than those of their parents, seed sets were similar to or of lower value than those of weedy rice parents and seed shattering characteristics were partially suppressed, but the survival of hybrids over winter in the field was similar to that of weedy rice parents. All F1, F2 and F3 hybrids had similar composite agronomic performance to weedy rice parents. CONCLUSION There was no significant decrease in the composite agronomic performance of any of the hybrids compared with weedy rice. This implies that gene flow from transgenic cultivated rice to weedy rice could occur under natural conditions.

Potential gene flow from transgenic rice (Oryza sativa L.) to different weedy rice (Oryza sativa f. spontanea) accessions based on reproductive compatibility

BACKGROUND The possibility of gene flow from transgenic crops to wild relatives may be affected by reproductive capacity between them. The potential gene flow from two transgenic rice lines containing the bar gene to five accessions of weedy rice (WR1-WR5) was determined through examination of reproductive compatibility under controlled pollination. RESULTS The pollen grain germination of two transgenic rice lines on the stigma of all weedy rice, rice pollen tube growth down the style and entry into the weedy rice ovary were similar to self-pollination in weedy rice. However, delayed double fertilisation and embryo abortion in crosses between WR2 and Y0003 were observed. Seed sets between transgenic rice lines and weedy rice varied from 8 to 76%. Although repeated pollination increased seed set significantly, the rank of the seed set between the weedy rice accessions and rice lines was not changed. The germination rates of F(1) hybrids were similar or greater compared with respective females. All F(1) plants expressed glufosinate resistance in the presence of glufosinate selection pressure. CONCLUSIONS The frequency of gene flow between different weedy rice accessions and transgenic herbicide-resistant rice may differ owing to different reproductive compatibility. This result suggests that, when wild relatives are selected as experimental materials for assessing the gene flow of transgenic rice, it is necessary to address the compatibility between transgenic rice and wild relatives.

Innate factors causing differences in gene flow frequency from transgenic rice to different weedy rice biotypes

BACKGROUND The compatibility and outcrossing rates between transgenic rice and weedy rice biotypes have been studied in some previous cases. However, few studies have addressed the reasons for these differences. The present study compared the compatibility and outcrossing rates between transgenic rice and selected weedy rice biotypes using manual and natural crossing experiments to elucidate the key innate factors causing the different outcrossing rates. RESULTS Hybrid seed sets from manual crossing between transgenic rice and weedy rice varied from 31.8 to 82.7%, which correlated directly with genetic compatibility. Moreover, the significant differences in the quantity of germinated donor pollens and pollen tubes entering the weedy rice ovule directly contributed to the different seed sets. The natural outcrossing rates varied from 0 to 6.66‰. The duration of flowering overlap was the key factor influencing natural outcrossing. Plant and panicle height also affected outcrossing success. CONCLUSION From this study, it is concluded that the likelihood of gene flow between transgenic rice and weedy rice biotypes is primarily determined by floral synchronisation and secondarily influenced by genetic compatibility and some morphological characteristics.

A comparative study of competitiveness between different genotypes of weedy rice (Oryza sativa) and cultivated rice

BACKGROUND Competition from weedy rice can cause serious yield losses to cultivated rice. However, key traits that facilitate competitiveness are still not well understood. To explore the mechanisms behind the strong growth and competitive ability, replacement series experiments were established with six genotypes of weedy rice from different regions and one cultivated rice cultivar. RESULTS (1) Weedy rice from southern China had the greatest impact on growth and yield of cultivated rice throughout the entire growing season. Weedy rice from the northeast was very competitive during the early vegetative stage while the competitive effects of eastern weedy rice were more detrimental at later crop-growth stages. (2) As the proportion of weedy rice increased, plant height, tillers, above-ground biomass, and yield of cultivated rice significantly declined; the crop always being at disadvantage regardless of proportion. (3) Weedy biotypes with greater diversity as estimated by their Shannon indexes were more detrimental to the growth and yield of cultivated rice. CONCLUSION Geographic origin (latitude) of weedy rice biotype, its mixture proportion under competition with the crop and its genetic diversity are determinant factors of the outcome of competition and the associated decline in the rice crop yield.

Occurrence of Glyphosate-Resistant Horseweed （Conyza canadensis） Population in China

Horseweed (Conyza canadensis), an invasive alien weed, is one of the main weeds in orchards in China. Although glyphosate has been used for control of horseweed and many other weeds in orchards for more than 25 years in China, a case of glyphosate-resistant horseweed has not been identified in orchard in China so far despite glyphosate-resistant horseweed cases have been reported in some other countries. Seeds of 25 horseweed populations were collected from different orchards with different glyphosate application history. Potted seedlings with 11-13-leaf growth stage were treated with glyphosate at 0.035, 0.07, 0.14, 0.28, 0.56, 1.12, 2.24, 4.48, and 8.96 kg a.i. ha -1 . The dosage dependence response curve of each population was constructed with Log-logistic dose response regression equations. The ED50 value of each population was calculated and compared with the susceptible population from China. Different populations had different relative glyphosate-resistant levels which increased with the number of years of glyphosate application. Two populations with the highest resistance levels, 8.28 and 7.95 times, were found in Ningbo, Zhejiang Province, China, where glyphosate was used for weed control in orchards twice each year for 15 yr. The two resistant populations accumulated approximately two to four times less shikimic acid than the two susceptible populations 48 h after glyphosate application.

Cytoplasmic-genetic male sterility gene provides direct evidence for some hybrid rice recently evolving into weedy rice

Weedy rice infests paddy fields worldwide at an alarmingly increasing rate. There is substantial evidence indicating that many weedy rice forms originated from or are closely related to cultivated rice. There is suspicion that the outbreak of weedy rice in China may be related to widely grown hybrid rice due to its heterosis and the diversity of its progeny, but this notion remains unsupported by direct evidence. We screened weedy rice accessions by both genetic and molecular marker tests for the cytoplasmic male sterility (CMS) genes (Wild abortive, WA, and Boro type, BT) most widely used in the production of indica and japonica three-line hybrid rice as a diagnostic trait of direct parenthood. Sixteen weedy rice accessions of the 358 tested (4.5%) contained the CMS-WA gene; none contained the CMS-BT gene. These 16 accessions represent weedy rices recently evolved from maternal hybrid rice derivatives, given the primarily maternal inheritance of this trait. Our results provide key direct evidence that hybrid rice can be involved in the evolution of some weedy rice accessions, but is not a primary factor in the recent outbreak of weedy rice in China.

A model of the relationship between weedy rice seed-bank dynamics and rice-crop infestation and damage in Jiangsu Province, China.

BACKGROUND A heavy infestation of weedy rice leading to no harvested rice has never been predicted in China due to a lack of knowledge about the weedy rice seed bank. We studied the seed-bank dynamics of weedy rice for three consecutive years and analyzed the relationship between seed-bank density and population density in order to predict future weedy rice infestations of direct-seeded rice at six sites along the Yangtze River in Jiangsu Province, China. RESULTS The seed-bank density of weedy rice in all six sites displayed an increasing trend with seasonal fluctuations. Weedy rice seeds found in the 0-10 cm soil layer contributed most to seedling emergence. An exponential curve expressed the relationship between cultivated rice yield loss and adult weedy rice density. Based on data collected during the weedy rice life-cycle, a semi-empirical mathematic model was developed that fits well with the experimental data in a way that could be used to predict seed-bank dynamics. CONCLUSIONS By integrating the semi-empirical model and the exponential curve, weedy rice infestation levels and crop losses can be predicted based on the seed-bank dynamics so that a practical control can be adopted before rice planting.

Enhanced photosynthesis endows seedling growth vigor contributing to the competitive dominance of weedy rice over cultivated rice.

BACKGROUND Weedy rice, as one of the worst paddy field weeds worldwide, bears vigorous seedlings and dominantly competes with cultivated rice causing serious crop yield losses. To elucidate the causes of its stronger seedling vigour endowing its dominant competition with cultivated rice, comparative studies on seedling growth characteristics were conducted among six weedy rice biotypes and the two indica and japonica cultivars Shanyou-63 (SY-63) and Zhendao-8 (ZD-8), respectively, in the greenhouse. RESULTS Weedy rice emerged 2 to 3 days earlier, rapidly grew 1.3-1.7 cm taller daily, produced more secondary adventitious roots and greater aboveground fresh biomass than cultivated rice. Moreover, weedy rice exhibited greater photosynthetic pigment content, net photosynthetic rate, stomatal conductance, intercellular CO2 concentration, transpiration rate, and chlorophyll fluorescence kinetic parameters. An enhanced overall photosynthetic activity in weedy rices was attributed to the combined action of a larger antenna, more active reaction centres and higher quantum yield for electron transfer beyond QA . CONCLUSIONS Enhanced photosynthesis of weedy rice at the seedling stage should be the main factor for leading to strong competitive dominance over cultivated rice.

Allele Types of Rc Gene of Weedy Rice from Jiangsu Province, China

Weedy rice (Oryza sativa f. spontanea), the predominant type of which has a red pericarp, seriously inhibits growth and yield of direct-seeded rice in Jiangsu Province, China. In this study, we randomly selected 10 weedy rice accessions from 10 plots in Jiangsu, and then sequenced the full lengths of their Rc genes (approximately 6.4 kb). In addition, we collected 166 different full-length Rc genes in the Oryza genus from the literature and from GenBank. A collinearity sequence analysis showed that the 10 weedy rice accessions from Jiangsu all had the same wild-type allele of the Rc gene. Single nucleotide polymorphisms indicated that the nucleotide polymorphisms (Π = 0.19) and the proportion of segregation sites (θw = 0.28) of the Rc genes in the 10 weedy rice accessions from Jiangsu were higher than those in 56 weedy rice accessions from USA (Π = 0.09 and θw = 0.07). Haplotype and phylogenetic analyses showed that the Rc genes of weedy rice accessions from Jiangsu were not revertants of the rc gene found in Asian cultivated rice (O. sativa) varieties with white pericarp. In addition, Rc gene sequences of the rice varieties Lvdao from Lianyungang, Jiangsu and Tangdao from Anhui were more similar to those of cultivated rice than to the weedy rice from Jiangsu. These findings support the continued quarantine of weedy rice and clarify the evolutionary mechanism of the red pericarp found in the weedy rice of Jiangsu.