Only few glufosinate-tolerant genes,such as phosphinothricin acetyltransferase(PAT)and bialaphos resistance(bar)identified from Streptomyces,are currently available for developing genetically modified rice in agricult...Only few glufosinate-tolerant genes,such as phosphinothricin acetyltransferase(PAT)and bialaphos resistance(bar)identified from Streptomyces,are currently available for developing genetically modified rice in agricultural application.Following the rapid development of genome editing technology,generation of novel glufosinate-tolerant gene resources through artificial evolution of endogenous genes is more promising and highly desirable in rice molecular breeding program.In this study,the endogenous Glutamine synthetase1(OsGS1)was artificially evolved by base-editing-mediated gene evolution(BEMGE)in rice cells to create novel alleles conferring glufosinate tolerance in rice germplasms.Two novel glufosinate-tolerant OsGS1 alleles(OsGS1-AVPS and OsGS1-+AF)and one reported tolerant allele(OsGS1-SGTA)were successfully identified from approximately 4200 independent hygromycin-tolerant calli.Germination assays and spray tests revealed that these three OsGS1 alleles confer glufosinate tolerance in rice.Furthermore,OsGS1-AVPS and OsGS1-SGTA were quickly deployed into the elite rice cultivar Nangeng 46 through precise base editing.Overall,our results demonstrate the feasibility of developing glufosinate-tolerant rice by editing an endogenous rice gene in molecular breeding programs.展开更多
The use of microbes and microbial products as bioherbicides has been studied for several decades, and combinations of bioherbicides and herbicides have been examined to discover possible synergistic interactions to im...The use of microbes and microbial products as bioherbicides has been studied for several decades, and combinations of bioherbicides and herbicides have been examined to discover possible synergistic interactions to improve weed control efficacy. Bioassays were conducted to assess possible interactions of the herbicide glufosinate [2-amino-4-(hydroxymethylphosphinyl) butanoic acid] and Colletotrichum truncatum (CT), a fungal bioherbicide to control hemp sesbania (Sesbania exaltata)]. Glufosinate acts as a glutamine synthetase (GS) inhibitor that causes elevated ammonia levels, but the mode of action of CT is unknown. GS has also been implicated in plant defense in certain plant-pathogen interactions. The effects of spray applications of glufosinate (1.0 mM) orbioherbicide (8.0 × 104 conidia ml-1), applied alone or in combination were monitored (88 h time-course) on seedling growth, GS activity and ammonia levels in hypocotyl tissues under controlled environmental conditions. Growth (elongation and fresh weight) and extractable GS activity were inhibited in tissues by glufosinate and glufosinate plus CT treatments as early as 16 h, but CT treatment did not cause substantial growth reduction or GS inhibition until after ~40 h. Generally, ammonia levels in hemp sesbania tissues under these various treatments were inversely correlated with GS activity. Localization of hemp sesbania GS activity on electrophoretic gels indicated a lack of activity after 30 h in glufosinate and glufosinate plus CT-treated tissue. Untreated control tissues contained much lower ammonia levels at 24, 64, and 88 h after treatment than treatments with CT, glufosinate or their combination. CT alone caused elevated ammonia levels only after 64 - 88 h. Glufosinate incorporated in agar at 0.25 mM to 2.0 mM, caused a 10% - 45% reduction of CT colony radial growth, compared to fungal growth on agar without glufosinate, and the herbicide also inhibited sporulation of CT. Although no synergistic interactions were found in the combinations of CT and glufosinate at the concentrations used, further insight on the biochemical action of CT and its interactions with this herbicide on hemp sesbania was achieved.展开更多
Palmer amaranth, sicklepod and pitted morningglory are the three most common and troublesome weeds in soybean in South Carolina. They exhibit very aggressive growth capabilities and if left uncontrolled in fields will...Palmer amaranth, sicklepod and pitted morningglory are the three most common and troublesome weeds in soybean in South Carolina. They exhibit very aggressive growth capabilities and if left uncontrolled in fields will cause significant reductions in soybean yields. Dicamba and 2,4-D herbicides are currently having a resurgence in usage due to the recent commercialization of soybean trait technologies with tolerance to these herbicides. Dicamba and 2,4-D when tank mixed with glufosinate and glyphosate may offer additional weed control to resistant weeds through the process of herbicide synergism. Greenhouse experiments were conducted in 2013 at Edisto Research and Education Center near Blackville, SC to evaluate the efficacy of glyphosate, glufosinate, dicamba and 2,4-D treatments alone and in combination on Palmer amaranth, sicklepod, and pitted morningglory at selected heights. Results suggested that glufosinate alone provided the overall best control for all 3 weed species. Glyphosate alone provided the lowest control of all 3 species at all heights. Synergism or improved sicklepod control was observed when glufosinate was tank mixed with dicamba. However, as sicklepod increased in height, glufosinate + 2,4-D or dicamba combination offered the best control compared to glufosinate alone (90% versus 86% in 20 cm plants and 87% versus 85% in 30 cm plant). In the 5 cm Palmer amaranth, decreased control was observed when glyphosate or glufosinate was tank mixed with 2,4-D. These experiments showed that glufosinate alone and/or in combination with 2,4-D or dicamba was the overall best treatment on the three broadleaf weed species.展开更多
In this study, the characteristic of three transformants named as B1C106-1, B1C106-2, and B1C106-3 were studied that carried three innate resistant genes Bph14, Bph15, and Xa23, and two enthetic resistant genes CrylCa...In this study, the characteristic of three transformants named as B1C106-1, B1C106-2, and B1C106-3 were studied that carried three innate resistant genes Bph14, Bph15, and Xa23, and two enthetic resistant genes CrylCa# and Bar. The five resistant genes were all verified by PCR and the two enthetic genes were identified in single copy insertion by Southern blot. At tillering stage, the CrylC and PAT （phosphinothricin acetyl transferase） protein contents in leaf, sheath, and stem of T2 generation were in the similar pattern： leaf〉stem〉sheath, and showed significant difference （P〈0.01） among three organs. The average contents of CrylC protein in plant of B1C106-1, B1C106-2, and BIC106-3 were 12.95, 6.57, and 11.30 μg g-1, respectively, and showed significant difference （P〈0.01） among them. However, the average contents of PAT in plant of B1C106-1, B1C106-2, and B1C106-3 were 28.54, 27.66, and 28.02 pg g-1, respectively, and there were no significant difference among three transformants. The glufosinate tolerable concentration of three transformants of T3 generation reached at least 6 g L-1, and the mortality of rice leaf rollers were above 97.4% in 5 days after being fed with fresh transformants＇ leaves. The CrylC protein toxicity was also assessed by silkworms, and the mortality of silkworms feeding mulberry leaves smeared with Cry1C protein extracts of leaves of B1 C106-1, BIC106-2, and B1 C106-3 were 90, 67.8, and 87.8%, respectively, that were positive correlation （r=0.993） with CrylC protein contents in plant of three transformants. The three transformants also maintained high resistance to brown planthopper and bacterial blight as the original version. The above results indicate the tetra-resistant rice germplasm was well-developed by pyramiding innate and enthetic resistant genes in an elite line to provide with resistances of glufosinate, rice leaf roller, brown planthopper, and bacterial blight.展开更多
Four field trials were conducted on a farm infested with glyphosate-resistant (GR) common ragweed during 2016 and 2017 to evaluate various postemergence (POST) herbicides for the control of GR common ragweed in GR cor...Four field trials were conducted on a farm infested with glyphosate-resistant (GR) common ragweed during 2016 and 2017 to evaluate various postemergence (POST) herbicides for the control of GR common ragweed in GR corn. Dicamba at 600 g·a.i.·ha-1, dicamba/diflufenzopyr at 200 g·a.i.·ha-1, dicamba/atrazine at 1500 g·a.i.·ha-1, topramezone + atrazine at 12.5 + 500 g·a.i.·ha-1, bromoxynil + atrazine at 280 + 1500 g·a.i.·ha-1, glufosinate at 500 g·a.i.·ha-1 and 2,4-D ester at 560 g·a.i.·ha-1 provided 58% to 85% control at 4 WAA and 49% to 88% control at 8 WAA. Other herbicides evaluated controlled GR common ragweed 9% to 41%. Common ragweed density was reduced 97%, 95%, 95% and 87% and shoot dry weight was reduced 93%, 95%, 94% and 90% with bromoxynil + atrazine, dicamba, glufosinate and topramezone + atrazine applied POST in GR corn, respectively. Results show that dicamba, bromoxynil + atrazine, topramezone + atrazine and glufosinate applied POST are the most efficacious herbicides among the herbicides evaluated for the control of GR common ragweed in GR corn.展开更多
The use of transgenic crops has grown significantly over the past couple of decades. Many agronomic crops produced today are tolerant to glyphosate. Glyphosate-tolerant crops were commercially introduced in 1996, and,...The use of transgenic crops has grown significantly over the past couple of decades. Many agronomic crops produced today are tolerant to glyphosate. Glyphosate-tolerant crops were commercially introduced in 1996, and, about nine years later, glyphosate-resistant Palmer amaranth was confirmed in Georgia. Glyphosate-resistant weeds arose from reliance on postemergence only glyphosate programs to control weeds in crops. New transgenic traits for glufosinate and 2,4-D choline have been developed, and evaluations of stacked traits and concurrent use of multiple herbicides have provided additional tools in the management of glyphosate-resistant weeds. Field experiments were conducted in 2012 and 2013 at the Edisto Research and Education Center near Blackville, SC, USA to determine the efficacy of 2,4-D-based herbicide programs in transgenic cotton tolerant to 2,4-D choline, glyphosate, and glufosinate. The treatments provided good to excellent Palmer amaranth and pitted morningglory control in 2012 and 2013. Seed cotton yields across treatments ranged from 0 to 2057 kg ha-1. This new trait technology package in cotton permits in-season postemergence use of 2,4-D choline, a herbicide mode of action not previously used postemergence in cotton, which can control resistant weeds, including Palmer amaranth if applied at the proper growth stage.展开更多
基金supported by grants from the Shenzhen Science and Technology Program(KQTD20180411143628272)Special Funds for Science Technology Innovation and Industrial Development of Shenzhen Dapeng New District(PT202101-02)+3 种基金the Hainan Yazhou Bay Seed Lab(B21HJ0215),the National Natural Science Foundation of China(32102294)the China National Postdoctoral Program for Innovative Talents(BX2020378)the China Postdoctoral Science Foundation(2020M672902)the Central Publicinterest Scientific Institution Basal Research Fund(Y2022PT24).
文摘Only few glufosinate-tolerant genes,such as phosphinothricin acetyltransferase(PAT)and bialaphos resistance(bar)identified from Streptomyces,are currently available for developing genetically modified rice in agricultural application.Following the rapid development of genome editing technology,generation of novel glufosinate-tolerant gene resources through artificial evolution of endogenous genes is more promising and highly desirable in rice molecular breeding program.In this study,the endogenous Glutamine synthetase1(OsGS1)was artificially evolved by base-editing-mediated gene evolution(BEMGE)in rice cells to create novel alleles conferring glufosinate tolerance in rice germplasms.Two novel glufosinate-tolerant OsGS1 alleles(OsGS1-AVPS and OsGS1-+AF)and one reported tolerant allele(OsGS1-SGTA)were successfully identified from approximately 4200 independent hygromycin-tolerant calli.Germination assays and spray tests revealed that these three OsGS1 alleles confer glufosinate tolerance in rice.Furthermore,OsGS1-AVPS and OsGS1-SGTA were quickly deployed into the elite rice cultivar Nangeng 46 through precise base editing.Overall,our results demonstrate the feasibility of developing glufosinate-tolerant rice by editing an endogenous rice gene in molecular breeding programs.
文摘The use of microbes and microbial products as bioherbicides has been studied for several decades, and combinations of bioherbicides and herbicides have been examined to discover possible synergistic interactions to improve weed control efficacy. Bioassays were conducted to assess possible interactions of the herbicide glufosinate [2-amino-4-(hydroxymethylphosphinyl) butanoic acid] and Colletotrichum truncatum (CT), a fungal bioherbicide to control hemp sesbania (Sesbania exaltata)]. Glufosinate acts as a glutamine synthetase (GS) inhibitor that causes elevated ammonia levels, but the mode of action of CT is unknown. GS has also been implicated in plant defense in certain plant-pathogen interactions. The effects of spray applications of glufosinate (1.0 mM) orbioherbicide (8.0 × 104 conidia ml-1), applied alone or in combination were monitored (88 h time-course) on seedling growth, GS activity and ammonia levels in hypocotyl tissues under controlled environmental conditions. Growth (elongation and fresh weight) and extractable GS activity were inhibited in tissues by glufosinate and glufosinate plus CT treatments as early as 16 h, but CT treatment did not cause substantial growth reduction or GS inhibition until after ~40 h. Generally, ammonia levels in hemp sesbania tissues under these various treatments were inversely correlated with GS activity. Localization of hemp sesbania GS activity on electrophoretic gels indicated a lack of activity after 30 h in glufosinate and glufosinate plus CT-treated tissue. Untreated control tissues contained much lower ammonia levels at 24, 64, and 88 h after treatment than treatments with CT, glufosinate or their combination. CT alone caused elevated ammonia levels only after 64 - 88 h. Glufosinate incorporated in agar at 0.25 mM to 2.0 mM, caused a 10% - 45% reduction of CT colony radial growth, compared to fungal growth on agar without glufosinate, and the herbicide also inhibited sporulation of CT. Although no synergistic interactions were found in the combinations of CT and glufosinate at the concentrations used, further insight on the biochemical action of CT and its interactions with this herbicide on hemp sesbania was achieved.
文摘Palmer amaranth, sicklepod and pitted morningglory are the three most common and troublesome weeds in soybean in South Carolina. They exhibit very aggressive growth capabilities and if left uncontrolled in fields will cause significant reductions in soybean yields. Dicamba and 2,4-D herbicides are currently having a resurgence in usage due to the recent commercialization of soybean trait technologies with tolerance to these herbicides. Dicamba and 2,4-D when tank mixed with glufosinate and glyphosate may offer additional weed control to resistant weeds through the process of herbicide synergism. Greenhouse experiments were conducted in 2013 at Edisto Research and Education Center near Blackville, SC to evaluate the efficacy of glyphosate, glufosinate, dicamba and 2,4-D treatments alone and in combination on Palmer amaranth, sicklepod, and pitted morningglory at selected heights. Results suggested that glufosinate alone provided the overall best control for all 3 weed species. Glyphosate alone provided the lowest control of all 3 species at all heights. Synergism or improved sicklepod control was observed when glufosinate was tank mixed with dicamba. However, as sicklepod increased in height, glufosinate + 2,4-D or dicamba combination offered the best control compared to glufosinate alone (90% versus 86% in 20 cm plants and 87% versus 85% in 30 cm plant). In the 5 cm Palmer amaranth, decreased control was observed when glyphosate or glufosinate was tank mixed with 2,4-D. These experiments showed that glufosinate alone and/or in combination with 2,4-D or dicamba was the overall best treatment on the three broadleaf weed species.
基金supported by the State Key Science and Technology Programme on Breeding of New Genetically Modified Organisms, China (2011ZX08001-003 and 2016ZX08001-003)the Open Research Fund of State Key Laboratory of Hybrid Rice (Hunan Hybrid Rice Research Center), China
文摘In this study, the characteristic of three transformants named as B1C106-1, B1C106-2, and B1C106-3 were studied that carried three innate resistant genes Bph14, Bph15, and Xa23, and two enthetic resistant genes CrylCa# and Bar. The five resistant genes were all verified by PCR and the two enthetic genes were identified in single copy insertion by Southern blot. At tillering stage, the CrylC and PAT （phosphinothricin acetyl transferase） protein contents in leaf, sheath, and stem of T2 generation were in the similar pattern： leaf〉stem〉sheath, and showed significant difference （P〈0.01） among three organs. The average contents of CrylC protein in plant of B1C106-1, B1C106-2, and BIC106-3 were 12.95, 6.57, and 11.30 μg g-1, respectively, and showed significant difference （P〈0.01） among them. However, the average contents of PAT in plant of B1C106-1, B1C106-2, and B1C106-3 were 28.54, 27.66, and 28.02 pg g-1, respectively, and there were no significant difference among three transformants. The glufosinate tolerable concentration of three transformants of T3 generation reached at least 6 g L-1, and the mortality of rice leaf rollers were above 97.4% in 5 days after being fed with fresh transformants＇ leaves. The CrylC protein toxicity was also assessed by silkworms, and the mortality of silkworms feeding mulberry leaves smeared with Cry1C protein extracts of leaves of B1 C106-1, BIC106-2, and B1 C106-3 were 90, 67.8, and 87.8%, respectively, that were positive correlation （r=0.993） with CrylC protein contents in plant of three transformants. The three transformants also maintained high resistance to brown planthopper and bacterial blight as the original version. The above results indicate the tetra-resistant rice germplasm was well-developed by pyramiding innate and enthetic resistant genes in an elite line to provide with resistances of glufosinate, rice leaf roller, brown planthopper, and bacterial blight.
文摘Four field trials were conducted on a farm infested with glyphosate-resistant (GR) common ragweed during 2016 and 2017 to evaluate various postemergence (POST) herbicides for the control of GR common ragweed in GR corn. Dicamba at 600 g·a.i.·ha-1, dicamba/diflufenzopyr at 200 g·a.i.·ha-1, dicamba/atrazine at 1500 g·a.i.·ha-1, topramezone + atrazine at 12.5 + 500 g·a.i.·ha-1, bromoxynil + atrazine at 280 + 1500 g·a.i.·ha-1, glufosinate at 500 g·a.i.·ha-1 and 2,4-D ester at 560 g·a.i.·ha-1 provided 58% to 85% control at 4 WAA and 49% to 88% control at 8 WAA. Other herbicides evaluated controlled GR common ragweed 9% to 41%. Common ragweed density was reduced 97%, 95%, 95% and 87% and shoot dry weight was reduced 93%, 95%, 94% and 90% with bromoxynil + atrazine, dicamba, glufosinate and topramezone + atrazine applied POST in GR corn, respectively. Results show that dicamba, bromoxynil + atrazine, topramezone + atrazine and glufosinate applied POST are the most efficacious herbicides among the herbicides evaluated for the control of GR common ragweed in GR corn.
文摘The use of transgenic crops has grown significantly over the past couple of decades. Many agronomic crops produced today are tolerant to glyphosate. Glyphosate-tolerant crops were commercially introduced in 1996, and, about nine years later, glyphosate-resistant Palmer amaranth was confirmed in Georgia. Glyphosate-resistant weeds arose from reliance on postemergence only glyphosate programs to control weeds in crops. New transgenic traits for glufosinate and 2,4-D choline have been developed, and evaluations of stacked traits and concurrent use of multiple herbicides have provided additional tools in the management of glyphosate-resistant weeds. Field experiments were conducted in 2012 and 2013 at the Edisto Research and Education Center near Blackville, SC, USA to determine the efficacy of 2,4-D-based herbicide programs in transgenic cotton tolerant to 2,4-D choline, glyphosate, and glufosinate. The treatments provided good to excellent Palmer amaranth and pitted morningglory control in 2012 and 2013. Seed cotton yields across treatments ranged from 0 to 2057 kg ha-1. This new trait technology package in cotton permits in-season postemergence use of 2,4-D choline, a herbicide mode of action not previously used postemergence in cotton, which can control resistant weeds, including Palmer amaranth if applied at the proper growth stage.
