TILLING(Targeting induced local lesions in genomes,定向诱导基因组局部突变技术)是一种高通量的等位变异创制和突变体快速鉴定技术,其实质是将传统的化学诱变方法和突变的高效筛选有效结合的反向遗传学研究方法。其技术原理是将传...TILLING(Targeting induced local lesions in genomes,定向诱导基因组局部突变技术)是一种高通量的等位变异创制和突变体快速鉴定技术,其实质是将传统的化学诱变方法和突变的高效筛选有效结合的反向遗传学研究方法。其技术原理是将传统的酶切技术与PCR技术相结合后采用红外双色荧光系统进行结果鉴定,从而筛选出相应的突变体。传统的TILLING技术主要用于筛选由人工诱导产生的突变体。Ecotilling技术由TILL-ING技术延伸而来,主要用于鉴定自然界中已经存在的突变体,其与传统的TILLING技术的区别主要为构建DNA池时略有差异。随着该项技术在拟南芥等模式植物中的成功应用,越来越多的人开始将其用于基因组较大的植物之中。本文对近年来TILLING技术在麦类作物中的应用进行了分析,并通过比较不同植物突变体库中的突变频率发现,经EMS处理的小麦等麦类作物突变体库中的突变频率显著高于其他植物,因此相信,TILLING技术将会作为一种常规手段在麦类作物尤其是普通小麦改良中得到越来越广泛的应用。展开更多
TILLING,即Targeting Induced Local Lesions IN Genomes(定向诱导基因组局部突变技术),是由美国Fred Hutchinson癌症研究中心Steven Henikoff领导的研究小组发展起来的,是一种全新的反向遗传学研究方法。TILLING技术借助高通量的检测手...TILLING,即Targeting Induced Local Lesions IN Genomes(定向诱导基因组局部突变技术),是由美国Fred Hutchinson癌症研究中心Steven Henikoff领导的研究小组发展起来的,是一种全新的反向遗传学研究方法。TILLING技术借助高通量的检测手段,快速有效地从由化学诱变剂(EMS)诱变过的突变群体中鉴定出点突变。目前,TILLING已被应用于多种生物的研究中。本文系统介绍了TILLING技术的基本原理、技术路线及其技术优势,同时列举了TILLING的应用实例,并展望了TILLING技术的应用前景。展开更多
Global food demand requires that soils be used intensively for agriculture, but how these soils are managed greatly impacts soil fluxes of carbon dioxide (CO2). Soil management practices can cause carbon to be either ...Global food demand requires that soils be used intensively for agriculture, but how these soils are managed greatly impacts soil fluxes of carbon dioxide (CO2). Soil management practices can cause carbon to be either sequestered or emitted, with corresponding uncertain influence on atmospheric CO2 concentrations. The situation is further complicated by the lack of CO2 flux measurements for African subsistence farms. For widespread application in remote areas, a simple experimental methodology is desired. As a first step, the present study investigated the use of Bowen Ratio Energy Balance (BREB) instrumentation to measure the energy balance and CO2 fluxes of two contrasting crop management systems, till and no-till, in the lowlands within the mountains of Lesotho. Two BREB micrometeorological systems were established on 100-m by 100-m sites, both planted with maize (Zea mays) but under either conventional (plow, disk-disk) or no-till soil mangement systems. The results demonstrate that with careful maintenance of the instruments by appropriately trained local personnel, the BREB approach offers substantial benefits in measuring real time changes in agroecosystem CO2 flux. The periods where the two treatments could be compared indicated greater CO2 sequestration over the no-till treatments during both the growing and non-growing seasons.展开更多
TILLING技术(Targeting Induced Local Lesions IN Genomes)经过十多年的快速发展,已经应用到多种植物点突变高通量筛选与基因功能研究中,检测方法也逐渐趋于多样化,直接测序、毛细管电泳、高分辨熔解曲线等检测方法均得到了较多的应用...TILLING技术(Targeting Induced Local Lesions IN Genomes)经过十多年的快速发展,已经应用到多种植物点突变高通量筛选与基因功能研究中,检测方法也逐渐趋于多样化,直接测序、毛细管电泳、高分辨熔解曲线等检测方法均得到了较多的应用。与此同时EcoTILLING、DeTILLING及iTILLING等相关技术的发展扩大了TILLING技术的应用范围。本文着重介绍了目前TILLING分析中用到的不同检测方法及相关生物信息学工具,详细评述了TILLING技术在模式植物、主要粮食作物、油料作物、蔬菜等其它作物中的应用与研究进展。展开更多
定向诱导基因组局部突变(targeting induced local lesions in genomes,TILLING)技术将化学诱变与高通量突变检测技术相结合,可高效、快速地从突变群体中鉴定出目标基因突变位点。本文在概述TILLING技术应用于水稻、小麦、玉米、大豆等...定向诱导基因组局部突变(targeting induced local lesions in genomes,TILLING)技术将化学诱变与高通量突变检测技术相结合,可高效、快速地从突变群体中鉴定出目标基因突变位点。本文在概述TILLING技术应用于水稻、小麦、玉米、大豆等作物突变研究现状的基础上,重点综述了TILLING分析群体构建与突变位点检测方法的技术改进与发展,探讨了TILLING技术目前存在的问题与前景。展开更多
定向诱导基因组局部突变(targetinginduced local lesions in genomes,TILLING)可快速、有效地鉴定和定向筛选突变,是一种全新的反向遗传学技术。介绍了TILLING的技术流程与核心,并对其在突变研究、反向遗传学及功能基因组学、SNP检测...定向诱导基因组局部突变(targetinginduced local lesions in genomes,TILLING)可快速、有效地鉴定和定向筛选突变,是一种全新的反向遗传学技术。介绍了TILLING的技术流程与核心,并对其在突变研究、反向遗传学及功能基因组学、SNP检测、资源创新与分析以及作物遗传改良等方面的应用进行了综述。展开更多
TILLING(Targeting Induced Local Lesions In Genomes)即定向诱导基因组局部突变技术,是近年发展起来的一种高通量检测点突变的反向遗传学方法。本研究从PCR模板量及酶切体系中CELI的酶量两个方面对家蚕TILLING技术的检测体系进行了优...TILLING(Targeting Induced Local Lesions In Genomes)即定向诱导基因组局部突变技术,是近年发展起来的一种高通量检测点突变的反向遗传学方法。本研究从PCR模板量及酶切体系中CELI的酶量两个方面对家蚕TILLING技术的检测体系进行了优化。结果表明:本实验在应用TILLING技术筛选家蚕突变体过程中,用于PCR扩增的家蚕基因组DNA模板最佳用量为20ng;CELI酶切的20μL反应体系中,加入的最适酶量为0.5μL(本实验室从芹菜中自提的CELI酶10倍稀释液)。本研究初步对TILLING技术应用于家蚕化学诱导突变检测体系进行了优化,为TILLING技术在家蚕功能基因组研究中的应用奠定了基础。展开更多
定向诱导基因组局部突变(Targeting Induced Local Lesions In Genomes,TILLING)技术自诞生以来在多种模式和非模式植物功能基因组学研究中得到了广泛应用,并获得了大量可用于基础研究的突变体材料和可应用于农作物遗传改良的新种质。...定向诱导基因组局部突变(Targeting Induced Local Lesions In Genomes,TILLING)技术自诞生以来在多种模式和非模式植物功能基因组学研究中得到了广泛应用,并获得了大量可用于基础研究的突变体材料和可应用于农作物遗传改良的新种质。这种高通量、低成本的反向遗传学研究方法为剑麻研究带来了新的机遇。目前国内外尚无TILLING 技术应用于剑麻研究的报道,而麻类作物亚麻中则已出现 TILLING 技术成功应用的报道。本文将概述 TILLING 技术的特点,对比其在亚麻中的应用,并结合剑麻 EMS 诱变群体构建策略和剑麻基因组学研究进展,综合探讨该技术在剑麻中的应用前景。展开更多
The tillage experiments for winter wheat were conducted on the slope farmland in Luoyang, Henan Province in the semihumid to arid loess plateau areas of North China. Different tillage methods including reduced tillage...The tillage experiments for winter wheat were conducted on the slope farmland in Luoyang, Henan Province in the semihumid to arid loess plateau areas of North China. Different tillage methods including reduced tillage(RT), no-till(NT), 2 crops/year(2C), subsoiling(SS), and conventional tillage(CT)were compared to determine the effects of tillage methods on soil water conservation, water availability, and wheat yields in a search for better farming systems in the areas. The NT and SS showed good effects on water conservation. The soil water storage increased 12 - 33 mm with NT and 9-24 mm with SS at the end of summer fallow periods. The soil evaporation with NT and SS decreased 7-8 mm and 34 - 36 mm during the fallow periods of 1999 and 2001, respectively. Evapotranspiration(ET)with NT and SS increased about 47 mm during wheat growth periods of 2000 to 2001. Treatment RT and 2C had low water storage and high water losses during the fallow periods. The winter wheat yields with conservation tillage practices were improved in the 2nd year, increased by 3, 5 and 8% with RT, NT and SS, respectively, compared with CT. The highest wheat yields were obtained with subsoiling, and the maximum economic benefits from no-till. All conservation tillage practices provided great benefits to saving energy and labors, reducing operation inputs, and increasing economic returns. No-till and subsoiling have shown promise in increasing water storage, reducing water loss, enhancing water availability, and saving energy, as well as increasing wheat yield.展开更多
文摘TILLING(Targeting induced local lesions in genomes,定向诱导基因组局部突变技术)是一种高通量的等位变异创制和突变体快速鉴定技术,其实质是将传统的化学诱变方法和突变的高效筛选有效结合的反向遗传学研究方法。其技术原理是将传统的酶切技术与PCR技术相结合后采用红外双色荧光系统进行结果鉴定,从而筛选出相应的突变体。传统的TILLING技术主要用于筛选由人工诱导产生的突变体。Ecotilling技术由TILL-ING技术延伸而来,主要用于鉴定自然界中已经存在的突变体,其与传统的TILLING技术的区别主要为构建DNA池时略有差异。随着该项技术在拟南芥等模式植物中的成功应用,越来越多的人开始将其用于基因组较大的植物之中。本文对近年来TILLING技术在麦类作物中的应用进行了分析,并通过比较不同植物突变体库中的突变频率发现,经EMS处理的小麦等麦类作物突变体库中的突变频率显著高于其他植物,因此相信,TILLING技术将会作为一种常规手段在麦类作物尤其是普通小麦改良中得到越来越广泛的应用。
文摘TILLING,即Targeting Induced Local Lesions IN Genomes(定向诱导基因组局部突变技术),是由美国Fred Hutchinson癌症研究中心Steven Henikoff领导的研究小组发展起来的,是一种全新的反向遗传学研究方法。TILLING技术借助高通量的检测手段,快速有效地从由化学诱变剂(EMS)诱变过的突变群体中鉴定出点突变。目前,TILLING已被应用于多种生物的研究中。本文系统介绍了TILLING技术的基本原理、技术路线及其技术优势,同时列举了TILLING的应用实例,并展望了TILLING技术的应用前景。
文摘Global food demand requires that soils be used intensively for agriculture, but how these soils are managed greatly impacts soil fluxes of carbon dioxide (CO2). Soil management practices can cause carbon to be either sequestered or emitted, with corresponding uncertain influence on atmospheric CO2 concentrations. The situation is further complicated by the lack of CO2 flux measurements for African subsistence farms. For widespread application in remote areas, a simple experimental methodology is desired. As a first step, the present study investigated the use of Bowen Ratio Energy Balance (BREB) instrumentation to measure the energy balance and CO2 fluxes of two contrasting crop management systems, till and no-till, in the lowlands within the mountains of Lesotho. Two BREB micrometeorological systems were established on 100-m by 100-m sites, both planted with maize (Zea mays) but under either conventional (plow, disk-disk) or no-till soil mangement systems. The results demonstrate that with careful maintenance of the instruments by appropriately trained local personnel, the BREB approach offers substantial benefits in measuring real time changes in agroecosystem CO2 flux. The periods where the two treatments could be compared indicated greater CO2 sequestration over the no-till treatments during both the growing and non-growing seasons.
文摘TILLING技术(Targeting Induced Local Lesions IN Genomes)经过十多年的快速发展,已经应用到多种植物点突变高通量筛选与基因功能研究中,检测方法也逐渐趋于多样化,直接测序、毛细管电泳、高分辨熔解曲线等检测方法均得到了较多的应用。与此同时EcoTILLING、DeTILLING及iTILLING等相关技术的发展扩大了TILLING技术的应用范围。本文着重介绍了目前TILLING分析中用到的不同检测方法及相关生物信息学工具,详细评述了TILLING技术在模式植物、主要粮食作物、油料作物、蔬菜等其它作物中的应用与研究进展。
文摘定向诱导基因组局部突变(targeting induced local lesions in genomes,TILLING)技术将化学诱变与高通量突变检测技术相结合,可高效、快速地从突变群体中鉴定出目标基因突变位点。本文在概述TILLING技术应用于水稻、小麦、玉米、大豆等作物突变研究现状的基础上,重点综述了TILLING分析群体构建与突变位点检测方法的技术改进与发展,探讨了TILLING技术目前存在的问题与前景。
基金This work was supported the National High Technology Research and Development Program of China (863 Program) (No. 2002AA2Z1003), Key Project of Chinese National Programs for Fundamental Research and Development (No.2005CB120801).
文摘定向诱导基因组局部突变(targetinginduced local lesions in genomes,TILLING)可快速、有效地鉴定和定向筛选突变,是一种全新的反向遗传学技术。介绍了TILLING的技术流程与核心,并对其在突变研究、反向遗传学及功能基因组学、SNP检测、资源创新与分析以及作物遗传改良等方面的应用进行了综述。
文摘TILLING(Targeting Induced Local Lesions In Genomes)即定向诱导基因组局部突变技术,是近年发展起来的一种高通量检测点突变的反向遗传学方法。本研究从PCR模板量及酶切体系中CELI的酶量两个方面对家蚕TILLING技术的检测体系进行了优化。结果表明:本实验在应用TILLING技术筛选家蚕突变体过程中,用于PCR扩增的家蚕基因组DNA模板最佳用量为20ng;CELI酶切的20μL反应体系中,加入的最适酶量为0.5μL(本实验室从芹菜中自提的CELI酶10倍稀释液)。本研究初步对TILLING技术应用于家蚕化学诱导突变检测体系进行了优化,为TILLING技术在家蚕功能基因组研究中的应用奠定了基础。
文摘The tillage experiments for winter wheat were conducted on the slope farmland in Luoyang, Henan Province in the semihumid to arid loess plateau areas of North China. Different tillage methods including reduced tillage(RT), no-till(NT), 2 crops/year(2C), subsoiling(SS), and conventional tillage(CT)were compared to determine the effects of tillage methods on soil water conservation, water availability, and wheat yields in a search for better farming systems in the areas. The NT and SS showed good effects on water conservation. The soil water storage increased 12 - 33 mm with NT and 9-24 mm with SS at the end of summer fallow periods. The soil evaporation with NT and SS decreased 7-8 mm and 34 - 36 mm during the fallow periods of 1999 and 2001, respectively. Evapotranspiration(ET)with NT and SS increased about 47 mm during wheat growth periods of 2000 to 2001. Treatment RT and 2C had low water storage and high water losses during the fallow periods. The winter wheat yields with conservation tillage practices were improved in the 2nd year, increased by 3, 5 and 8% with RT, NT and SS, respectively, compared with CT. The highest wheat yields were obtained with subsoiling, and the maximum economic benefits from no-till. All conservation tillage practices provided great benefits to saving energy and labors, reducing operation inputs, and increasing economic returns. No-till and subsoiling have shown promise in increasing water storage, reducing water loss, enhancing water availability, and saving energy, as well as increasing wheat yield.
