CN104642104A - Method for constructing alfalfa EMS mutant library - Google Patents
Method for constructing alfalfa EMS mutant library Download PDFInfo
- Publication number
- CN104642104A CN104642104A CN201510070291.6A CN201510070291A CN104642104A CN 104642104 A CN104642104 A CN 104642104A CN 201510070291 A CN201510070291 A CN 201510070291A CN 104642104 A CN104642104 A CN 104642104A
- Authority
- CN
- China
- Prior art keywords
- ems
- alfalfa
- seeds
- solution
- distilled water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 title claims abstract description 52
- 241000219823 Medicago Species 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 28
- 239000000463 material Substances 0.000 claims abstract description 12
- 238000012216 screening Methods 0.000 claims abstract description 8
- 238000011160 research Methods 0.000 claims abstract description 6
- 238000011161 development Methods 0.000 claims abstract description 4
- PLUBXMRUUVWRLT-UHFFFAOYSA-N Ethyl methanesulfonate Chemical compound CCOS(C)(=O)=O PLUBXMRUUVWRLT-UHFFFAOYSA-N 0.000 claims description 74
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- 239000000243 solution Substances 0.000 claims description 28
- 239000012153 distilled water Substances 0.000 claims description 22
- 238000009331 sowing Methods 0.000 claims description 17
- 235000015097 nutrients Nutrition 0.000 claims description 14
- 239000002689 soil Substances 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 7
- 241000196324 Embryophyta Species 0.000 claims description 6
- 239000000411 inducer Substances 0.000 claims description 6
- 239000008055 phosphate buffer solution Substances 0.000 claims description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 5
- 238000010276 construction Methods 0.000 claims description 4
- 238000003306 harvesting Methods 0.000 claims description 4
- 239000002361 compost Substances 0.000 claims description 3
- 238000009826 distribution Methods 0.000 claims description 3
- 239000003337 fertilizer Substances 0.000 claims description 3
- 239000003415 peat Substances 0.000 claims description 3
- 239000010451 perlite Substances 0.000 claims description 3
- 235000019362 perlite Nutrition 0.000 claims description 3
- 238000009394 selective breeding Methods 0.000 claims description 3
- 239000010455 vermiculite Substances 0.000 claims description 3
- 229910052902 vermiculite Inorganic materials 0.000 claims description 3
- 235000019354 vermiculite Nutrition 0.000 claims description 3
- 238000005507 spraying Methods 0.000 claims description 2
- 238000009395 breeding Methods 0.000 abstract description 5
- 230000001488 breeding effect Effects 0.000 abstract description 5
- 230000009418 agronomic effect Effects 0.000 abstract description 3
- 230000002068 genetic effect Effects 0.000 abstract description 3
- 239000007788 liquid Substances 0.000 description 6
- 240000004658 Medicago sativa Species 0.000 description 5
- 239000002962 chemical mutagen Substances 0.000 description 3
- 231100000350 mutagenesis Toxicity 0.000 description 3
- 229920006395 saturated elastomer Polymers 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 244000025254 Cannabis sativa Species 0.000 description 2
- 241000219793 Trifolium Species 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 241000219195 Arabidopsis thaliana Species 0.000 description 1
- 101710138460 Leaf protein Proteins 0.000 description 1
- 235000010624 Medicago sativa Nutrition 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 238000012214 genetic breeding Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明公开了一种紫花苜蓿EMS突变体库构建的方法,是以现有的紫花苜蓿栽培品种为材料,经一定浓度的EMS溶液处理一定时间,利用苗生长发育的各阶段进行突变体鉴定、筛选而获得该品种紫花苜蓿EMS突变体库的方法。本发明的方法具有操作简便、高效稳定、成本低等特点。利用该方法,可通过生物学性状和农艺性状的鉴定、筛选,得到用于该品种遗传改良的育种材料,同时,为紫花苜蓿的正向遗传学和功能基因组学分析提供丰富的研究材料。The invention discloses a method for constructing an alfalfa EMS mutant library. The existing alfalfa cultivars are used as materials, treated with a certain concentration of EMS solution for a certain period of time, and the mutants are identified at various stages of seedling growth and development. A method for obtaining the EMS mutant library of the variety alfalfa by screening. The method of the invention has the characteristics of simple and convenient operation, high efficiency and stability, low cost and the like. Using this method, the breeding material for the genetic improvement of the variety can be obtained through the identification and screening of biological and agronomic characters, and at the same time, it can provide abundant research materials for the forward genetics and functional genomics analysis of alfalfa.
Description
技术领域technical field
本发明涉及一种植物突变体库构建的方法,尤其涉及一种紫花苜蓿甲基磺酸乙酯(EMS)突变体库构建的方法。The invention relates to a method for constructing a plant mutant library, in particular to a method for constructing an alfalfa ethyl methanesulfonate (EMS) mutant library.
背景技术Background technique
紫花苜蓿(Medicago sativa L.)具有适应性广、高产优质、改良土壤、经济效益高、生态功能齐全的特性,素有“牧草之王”的美称。紫花苜蓿富含蛋白质、多种维生素和矿物质,是多种家禽喜食的优质蛋白质饲草和能量饲料,它既可用于青饲,同时也被用来开发研制成产品,包括草捆、草粉、草块、草颗粒和浓缩叶蛋白等。紫花苜蓿在畜牧业中发挥了决定性的作用,已成为我国畜牧业种植结构调整的首选饲料作物。另外,紫花苜蓿适应性极强、分枝多,不仅能固氮改土、保持水土,还对土壤的改良和生态环境的改善有重要的作用。Alfalfa (Medicago sativa L.) has the characteristics of wide adaptability, high yield and high quality, soil improvement, high economic benefits, and complete ecological functions. It is known as the "king of pastures". Alfalfa is rich in protein, vitamins and minerals. It is a high-quality protein forage and energy feed that many poultry like to eat. Powder, grass cubes, grass granules and leaf protein concentrate, etc. Alfalfa has played a decisive role in animal husbandry and has become the first choice feed crop for the adjustment of planting structure of animal husbandry in my country. In addition, alfalfa has strong adaptability and many branches. It can not only fix nitrogen to improve soil and maintain water and soil, but also play an important role in improving soil and ecological environment.
构建饱和突变体库是进行植物功能基因组学与遗传育种研究的直接而有效的方法之一。目前已在模式植物拟南芥、水稻等作物中构建了饱和突变体库并进行了相关的功能基因组学研究,获得了一批有价值的创新种质资源。若构建紫花苜蓿EMS突变体库之后,可通过生物学性状和农艺性状的鉴定、筛选,得到用于该品种遗传改良的育种材料,同时,为紫花苜蓿的正向遗传学和功能基因组学分析提供丰富的研究材料。Constructing a saturated mutant library is one of the most direct and effective methods for the study of plant functional genomics and genetic breeding. At present, saturated mutant libraries have been constructed in model plants such as Arabidopsis thaliana and rice, and related functional genomics studies have been carried out, and a batch of valuable innovative germplasm resources have been obtained. After the construction of the alfalfa EMS mutant library, the identification and screening of biological and agronomic traits can be used to obtain breeding materials for the genetic improvement of the variety, and at the same time, provide a basis for the forward genetics and functional genomics analysis of alfalfa. Rich research material.
化学诱变育种是人为的利用化学诱变剂,诱发作物发生突变,通过不断的筛选和鉴定,直接或间接地培育出能在直接利用于生产中的作物新品种,因而化学诱变是目前为止迅速发展农作物育种的手段。在众多的化学诱变剂中,甲基磺酸乙酯(Ethyl methanesulphonate,EMS)是应用最广泛、操作简便、高效、稳定的化学诱变剂。但在公知的甲基磺酸乙酯(EMS)诱变方法中针对紫花苜蓿EMS处理并构建EMS突变体库的优化方法还未见报道。Chemical mutation breeding is the artificial use of chemical mutagens to induce mutations in crops. Through continuous screening and identification, new crop varieties that can be directly or indirectly used in production are bred directly or indirectly. Rapidly develop means of crop breeding. Among many chemical mutagens, ethyl methanesulphonate (EMS) is the most widely used, easy to operate, efficient and stable chemical mutagen. However, in the well-known ethyl methanesulfonate (EMS) mutagenesis method, an optimized method for EMS treatment of alfalfa and construction of an EMS mutant library has not been reported.
发明内容Contents of the invention
针对现有技术的不足,本发明主要解决的问题是提供一种紫花苜蓿EMS突变体库构建的方法。Aiming at the deficiencies of the prior art, the main problem to be solved by the present invention is to provide a method for constructing an alfalfa EMS mutant library.
本发明所述紫花苜蓿EMS突变体库构建的方法,步骤是:The method for constructing the alfalfa EMS mutant library of the present invention, the steps are:
1)种子准备:选择饱满度高的紫花苜蓿种子,并按设定量称取该种子作为诱导剂处理材料;1) Seed preparation: select alfalfa seeds with high plumpness, and weigh the seeds as inducer treatment materials according to the set amount;
2)甲基磺酸乙酯(EMS)溶液准备:配制0.1mol/L、pH5.8的磷酸缓冲液并将诱导剂甲基磺酸乙酯(EMS)溶于该磷酸缓冲液中,制得甲基磺酸乙酯(EMS)溶液;2) Preparation of ethyl methanesulfonate (EMS) solution: prepare a 0.1mol/L, pH5.8 phosphate buffer solution and dissolve the inducer ethyl methanesulfonate (EMS) in the phosphate buffer solution to obtain Ethyl methanesulfonate (EMS) solution;
其中,所述pH5.8的0.1mol/L磷酸缓冲液的配制方法是:将1mol/L的Na2HPO4溶液定为A液;将1mol/L的NaH2PO4溶液定为B液。分别取A液92.1mL和B液7.9mL,加入到量筒中,定容至1L。Wherein, the preparation method of the 0.1 mol/L phosphate buffer solution with pH 5.8 is as follows: 1 mol/L Na 2 HPO 4 solution is defined as liquid A; 1 mol/L NaH 2 PO 4 solution is defined as B liquid. Take 92.1mL of liquid A and 7.9mL of liquid B respectively, add them to the graduated cylinder, and make the volume to 1L.
3)处理种子:用甲基磺酸乙酯(EMS)溶液处理紫花苜蓿种子;3) Treatment of seeds: treatment of alfalfa seeds with ethyl methanesulfonate (EMS) solution;
4)播种:对处理之后的紫花苜蓿种子分别编号,并播种于含育苗营养土的营养钵中育苗;4) sowing: numbering the alfalfa seeds after the treatment respectively, and sowing them in the nutrient bowl containing the nutrient soil for seedling cultivation;
5)移栽:待紫花苜蓿苗长出四片真叶后,将其移入起垄好的大田;苗生长期间加强肥水管理;5) Transplanting: After the alfalfa seedlings grow four true leaves, move them into the well-rided field; strengthen fertilizer and water management during the growth period of the seedlings;
6)收种:在苗生长发育的各阶段进行突变体鉴定、筛选、编号,单株收种并记载相关性状;6) Seed harvesting: Mutant identification, screening, and numbering are carried out at each stage of seedling growth and development, and seed is harvested from a single plant and related traits are recorded;
7)将获得各突变体分别编号并记载性状,建立紫花苜蓿甲基磺酸乙酯(EMS)突变体库,用于后续的紫花苜蓿选择育种和基因功能组学研究;7) The obtained mutants will be numbered and their traits recorded, and the alfalfa ethyl methanesulfonate (EMS) mutant library will be established for subsequent alfalfa selective breeding and gene functional omics research;
其特征在于:It is characterized by:
步骤2)所述甲基磺酸乙酯(EMS)溶液浓度以重量百分比计为1.6%;Step 2) the concentration of the ethyl methanesulfonate (EMS) solution is 1.6% by weight percent;
步骤3)所述处理种子的方法是:现将紫花苜蓿种子用蒸馏水浸泡24±4小时,然后去除蒸馏水并用步骤2)所述EMS溶液浸泡处理8±2小时,处理期间将含种子的容器置于摇床上以60±5rpm的转速轻摇;EMS溶液处理结束后,倒掉EMS溶液,将蒸馏水加入含种子的容器中并浸没种子,将该容器置于摇床上以60±5rpm的转速轻摇,每次30分钟,之后倒掉蒸馏水,再向该容器中加入蒸馏水并浸没种子,置于摇床上以60±5rpm的转速轻摇,每次30分钟,之后如此重复用蒸馏水冲洗EMS处理后的种子3-4次直至去除种子中残留的EMS;Step 3) The method for treating the seeds is: now soak the alfalfa seeds with distilled water for 24 ± 4 hours, then remove the distilled water and use the EMS solution described in step 2) to soak for 8 ± 2 hours, and place the container containing the seeds during the treatment. Gently shake at a speed of 60±5rpm on a shaker; after the EMS solution treatment, pour off the EMS solution, add distilled water into the container containing the seeds and submerge the seeds, place the container on a shaker at a speed of 60±5rpm and shake gently , 30 minutes each time, then pour out the distilled water, then add distilled water to the container and immerse the seeds, place it on a shaker at a speed of 60±5rpm, shake it gently for 30 minutes each time, and then repeatedly rinse the EMS-treated seeds with distilled water Seed 3-4 times until the residual EMS in the seed is removed;
步骤4)所述播种方法是:将处理后的种子播于直径6厘米,高10厘米的无纺布育苗营养钵中,人工穴播,播深1-2厘米;一天喷水3-4次;其中所述育苗营养土的组分配比以重量份计为:蛭石:泥炭:珍珠岩:腐熟有机堆肥=1:2:2:3。Step 4) The sowing method is: sowing the treated seeds in a 6 cm in diameter and 10 cm high non-woven seedling raising nutrient bowl, artificial hole sowing, sowing 1-2 cm deep; spraying water 3-4 times a day; Wherein the component distribution ratio of the nutrient soil for raising seedlings is calculated in parts by weight: vermiculite: peat: perlite: decomposed organic compost=1:2:2:3.
上述紫花苜蓿EMS突变体库构建的方法中:步骤3)所述处理种子的方法优选是:现将紫花苜蓿种子用蒸馏水浸泡24小时,然后去除蒸馏水并用步骤2)所述EMS溶液浸泡处理8小时,其中EMS溶液浓度以重量百分比计为1.6%。In the method for constructing the above-mentioned alfalfa EMS mutant library: the method for treating seeds described in step 3) is preferably: the alfalfa seeds are now soaked in distilled water for 24 hours, then the distilled water is removed and soaked in the EMS solution described in step 2) for 8 hours , wherein the concentration of the EMS solution is 1.6% by weight.
本发明公开了一种紫花苜蓿EMS突变体库构建的方法,以现有的紫花苜蓿栽培品种为材料,是经一定浓度的EMS溶液处理一定时间而获得该品种紫花苜蓿EMS突变体库的方法。本发明的方法具有操作简便、高效稳定、成本低等特点。利用该方法,可通过生物学性状和农艺性状的鉴定、筛选,得到用于该品种遗传改良的育种材料,同时,为紫花苜蓿的正向遗传学和功能基因组学分析提供丰富的研究材料。The invention discloses a method for constructing an alfalfa EMS mutant library. The existing alfalfa cultivars are used as materials, and the method is to obtain the alfalfa EMS mutant library of the variety after being treated with a certain concentration of EMS solution for a certain period of time. The method of the invention has the characteristics of simple and convenient operation, high efficiency and stability, low cost and the like. Using this method, the breeding material for the genetic improvement of the variety can be obtained through the identification and screening of biological and agronomic characters, and at the same time, it can provide abundant research materials for the forward genetics and functional genomics analysis of alfalfa.
具体实施方式Detailed ways
下面结合实施例对本发明做进一步详细说明。The present invention will be described in further detail below in conjunction with the examples.
实施例1:以紫花苜蓿栽培品种中苜一号为实验材料进行说明。Example 1: The alfalfa cultivar Zhonglu No. 1 is used as the experimental material for illustration.
紫花苜蓿(苜一号)EMS突变体库构建的方法,步骤是:The method for the construction of alfalfa (clover No. 1) EMS mutant library, the steps are:
1)种子准备:人工挑选饱满度高的苜一号紫花苜蓿种子作为诱导剂处理材料。为保证得到紫花苜蓿EMS饱和突变体库,严格称量320克的紫花苜蓿种子。1) Seed preparation: Alfalfa No. 1 alfalfa seeds with high plumpness were manually selected as inducer treatment materials. In order to ensure that the alfalfa EMS saturated mutant library was obtained, 320 grams of alfalfa seeds were strictly weighed.
2)EMS溶液准备:将1mol/L的Na2HPO4溶液定为A液;将1mol/L的NaH2PO4溶液定为B液。分别取A液92.1mL和B液7.9mL,加入到量筒中,定容至1L,得pH5.8的0.1mol/L磷酸缓冲液。将诱导剂甲基磺酸乙酯(EMS)溶于该磷酸缓冲液中,制得不同浓度的甲基磺酸乙酯(EMS)溶液。2) EMS solution preparation: 1 mol/L Na 2 HPO 4 solution is defined as A solution; 1 mol/L NaH 2 PO 4 solution is defined as B solution. Take 92.1mL of liquid A and 7.9mL of liquid B respectively, add them to a graduated cylinder, and dilute to 1L to obtain a 0.1mol/L phosphate buffer solution with a pH of 5.8. The inducer ethyl methanesulfonate (EMS) was dissolved in the phosphate buffer to prepare ethyl methanesulfonate (EMS) solutions with different concentrations.
其中,EMS浓度以重量百分比计分别为0、0.4%、0.8%、1.2%、1.6%、2.0%、2.4%、2.8%、3.2%。Wherein, the EMS concentration is respectively 0, 0.4%, 0.8%, 1.2%, 1.6%, 2.0%, 2.4%, 2.8%, 3.2% in weight percentage.
3)处理种子:将紫花苜蓿(苜一号)种子分组,分别先置于1L三角瓶中,用蒸馏水浸泡24小时。去除蒸馏水后,分别加入上述各浓度的EMS溶液对种子浸泡处理8小时、12小时、18小时,处理期间将含种子的容器置于摇床上以60rpm的转速轻摇;EMS溶液处理结束后,倒掉EMS溶液,将蒸馏水加入含种子的容器中并浸没种子,将该容器置于摇床上以60rpm的转速轻摇,每次30分钟,之后倒掉蒸馏水,再向该容器中加入蒸馏水并浸没种子,置于摇床上以60rpm的转速轻摇,每次30分钟,之后如此重复用蒸馏水冲洗EMS处理后的种子3-4次直至去除种子中残留的EMS。3) Seed treatment: alfalfa (Clover No. 1) seeds were divided into groups, placed in 1L Erlenmeyer flasks, and soaked in distilled water for 24 hours. After removing the distilled water, add the above-mentioned EMS solutions of various concentrations to soak the seeds for 8 hours, 12 hours, and 18 hours. During the treatment, place the container containing the seeds on a shaking table and shake gently at a speed of 60 rpm; Remove the EMS solution, add distilled water to the container containing the seeds and immerse the seeds, place the container on a shaker at a speed of 60 rpm for 30 minutes each time, then pour off the distilled water, then add distilled water to the container and immerse the seeds , placed on a shaker at a speed of 60 rpm for 30 minutes each time, and then repeatedly rinsed the EMS-treated seeds with distilled water for 3-4 times until the residual EMS in the seeds was removed.
4)播种:对处理之后的各组紫花苜蓿种子分别编号,并播种于含育苗营养土的营养钵中育苗;其中,所述播种方法是:将处理后的种子播于直径6厘米,高10厘米的无纺布育苗营养钵中,人工穴播,播深1-2厘米;营养钵置于40厘米×60厘米×10厘米营养托盘中,一天喷水3-4次;所述育苗营养土的组分配比以重量份计为:蛭石:泥炭:珍珠岩:腐熟有机堆肥=1:2:2:3。4) sowing: each group of alfalfa seeds after treatment was numbered respectively, and sowed in a nutrient pot containing seedling-raising nutrient soil for seedling cultivation; wherein, the sowing method was: sowing the treated seeds in a diameter of 6 cm, a height of 10 In a non-woven cloth seedling raising nutrient bowl of 1 cm, artificial hole sowing, sowing depth of 1-2 cm; the nutrient pot is placed in a nutrient tray of 40 cm × 60 cm × 10 cm, sprayed with water 3-4 times a day; the seedling nutrient soil The component distribution ratio is calculated in parts by weight: vermiculite: peat: perlite: decomposed organic compost = 1:2:2:3.
5)移栽:待紫花苜蓿苗长出四片真叶后,将其移入起垄好的大田;苗生长期间加强肥水管理。5) Transplanting: After the alfalfa seedlings grow four true leaves, move them into the well-ridden field; strengthen the management of fertilizer and water during the growth of the seedlings.
6)收种:在苗子生长发育的各阶段进行突变体鉴定、筛选、编号,单株收种并记载相关性状。6) Seed harvesting: Mutant identification, screening, numbering are carried out at each stage of seedling growth and development, single plant harvesting and related traits are recorded.
7)将获得的各突变体分别编号并记载性状,建立紫花苜蓿甲基磺酸乙酯(EMS)突变体库,用于后续的紫花苜蓿选择育种和基因功能组学研究。7) The obtained mutants were numbered and their traits were recorded, and an alfalfa ethyl methanesulfonate (EMS) mutant library was established for subsequent selective breeding of alfalfa and gene functional omics research.
上述试验中所用EMS浓度分别为0、0.4%、0.8%、1.2%、1.6%、2.0%、2.4%、2.8%、3.2%,处理紫花苜蓿中苜一号种子的时间梯度为8小时、12小时、18小时,每个处理重复3次,每个重复处理的种子为200粒。The EMS concentrations used were 0, 0.4%, 0.8%, 1.2%, 1.6%, 2.0%, 2.4%, 2.8%, 3.2% respectively in the above-mentioned test, and the time gradient for the treatment of No. 1 seed in alfalfa was 8 hours, 12 hours. Hour, 18 hours, each treatment was repeated 3 times, and the seeds of each repeated treatment were 200 seeds.
由上述EMS处理的浓度、时间及实验结果统计分析后得出:选择诱导紫花苜蓿中苜一号种子产生突变的甲基磺酸乙酯(EMS)半致死剂量浓度1.6%和8小时的处理时间,故本发明将其确定为构建紫花苜蓿EMS突变体库的最佳处理条件(见表1)。Draw after the concentration, time and experimental result statistical analysis of above-mentioned EMS treatment: select the half-lethal dose concentration of ethyl methanesulfonate (EMS) 1.6% and the processing time of 8 hours to produce mutation in alfalfa No. 1 seed , so the present invention determines it as the optimal processing condition for constructing alfalfa EMS mutant library (see Table 1).
表1:不同EMS浓度、不同时间梯度对中苜一号种子成苗率的影响(%)Table 1: Effects of different EMS concentrations and different time gradients on the seedling rate of Zhonglu No. 1 seeds (%)
Claims (2)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201510070291.6A CN104642104A (en) | 2015-02-10 | 2015-02-10 | Method for constructing alfalfa EMS mutant library |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201510070291.6A CN104642104A (en) | 2015-02-10 | 2015-02-10 | Method for constructing alfalfa EMS mutant library |
Publications (1)
Publication Number | Publication Date |
---|---|
CN104642104A true CN104642104A (en) | 2015-05-27 |
Family
ID=53234166
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201510070291.6A Pending CN104642104A (en) | 2015-02-10 | 2015-02-10 | Method for constructing alfalfa EMS mutant library |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN104642104A (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108419669A (en) * | 2018-02-28 | 2018-08-21 | 山东农业大学 | A kind of EMS directed mutagenesis, the method and device for screening salt tolerant alfalfa |
CN108617502A (en) * | 2018-04-26 | 2018-10-09 | 贵州省蚕业研究所 | A method of building capsicum mutant library using ethylmethane sulfonate |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008017718A (en) * | 2006-07-10 | 2008-01-31 | Institute Of Physical & Chemical Research | Photosynthetic organisms that green in a completely dark place |
WO2010042865A2 (en) * | 2008-10-10 | 2010-04-15 | Arcadia Biosciences, Inc. | Non-transgenic tomato varieties having increased shelf life post-harvest |
CN102812803A (en) * | 2012-07-02 | 2012-12-12 | 青岛市农业科学研究院 | Method for constructing pea EMS mutant library |
CN102835300A (en) * | 2011-06-23 | 2012-12-26 | 匡贤彦 | Method for efficiently obtaining tiller mutant through W64A pollen mutagenesis by using EMS |
CN103070068A (en) * | 2013-02-21 | 2013-05-01 | 江苏省农业科学院 | Herbicide-resistant rape directive breeding method based on acetolactate synthase (ALS) target esterase |
CN103299900A (en) * | 2013-05-03 | 2013-09-18 | 山西省农业科学院小麦研究所 | Wheat mutation breeding method |
-
2015
- 2015-02-10 CN CN201510070291.6A patent/CN104642104A/en active Pending
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008017718A (en) * | 2006-07-10 | 2008-01-31 | Institute Of Physical & Chemical Research | Photosynthetic organisms that green in a completely dark place |
WO2010042865A2 (en) * | 2008-10-10 | 2010-04-15 | Arcadia Biosciences, Inc. | Non-transgenic tomato varieties having increased shelf life post-harvest |
CN102835300A (en) * | 2011-06-23 | 2012-12-26 | 匡贤彦 | Method for efficiently obtaining tiller mutant through W64A pollen mutagenesis by using EMS |
CN102812803A (en) * | 2012-07-02 | 2012-12-12 | 青岛市农业科学研究院 | Method for constructing pea EMS mutant library |
CN103070068A (en) * | 2013-02-21 | 2013-05-01 | 江苏省农业科学院 | Herbicide-resistant rape directive breeding method based on acetolactate synthase (ALS) target esterase |
CN103299900A (en) * | 2013-05-03 | 2013-09-18 | 山西省农业科学院小麦研究所 | Wheat mutation breeding method |
Non-Patent Citations (2)
Title |
---|
R. VARMA PENMETSA AND DOUGLAS R. COOK: ""Production and Characterization of Diverse Developmental Mutants of Medicago truncatula1"", 《PLANT PHYSIOLOGY》 * |
夏曾润等: ""紫花苜蓿EMS突变体库的构建和形态学性状鉴定"", 《草业学报》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108419669A (en) * | 2018-02-28 | 2018-08-21 | 山东农业大学 | A kind of EMS directed mutagenesis, the method and device for screening salt tolerant alfalfa |
CN108419669B (en) * | 2018-02-28 | 2023-11-17 | 山东农业大学 | Method and device for EMS (enhanced message service) directional mutagenesis and screening salt-tolerant alfalfa |
CN108617502A (en) * | 2018-04-26 | 2018-10-09 | 贵州省蚕业研究所 | A method of building capsicum mutant library using ethylmethane sulfonate |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101817708B (en) | Plant source seed soaking agent and preparation method thereof | |
CN108929135B (en) | Root-promoting seedling-raising suspension type liquid fertilizer and preparation method and application thereof | |
CN106857211A (en) | A kind of authentication method of Fresh edible soybean time of infertility salt tolerance | |
CN102835297A (en) | Identification method of salt tolerance of barley | |
CN106258956A (en) | A kind of Herba Dendrobii pedigree breeding and field implantation method | |
CN108849491A (en) | A method of three-coloured amaranth seed is handled using ethylmethane sulfonate | |
Chakraborty et al. | Standarization of the appropriate doses of GA3 and Ag-nanoparticle in green gram for quality seed production | |
CN104642104A (en) | Method for constructing alfalfa EMS mutant library | |
CN117303946B (en) | Composite microalgae fertilizer capable of improving salt stress resistance of crops | |
Akon et al. | Influence of organic manures on the growth of diabetes preventive medicinal plant Gynura, Gynura procumbens | |
CN104012382A (en) | Large-scale seedling culturing technology for rhododendron calophytum Franch | |
Hossain et al. | Evaluation of mungbean (Vigna radiata (L.) Wilczek) genotypes on the basis of photosynthesis and dry matter accumulation | |
CN108094194A (en) | A kind of selection of richness potassium tobacco bred | |
Vasin et al. | Spring wheat productivity when using Megamix liquid fertilizers | |
CN106718864A (en) | A kind of method of the germplasm innovation of tawny daylily | |
Hongthong et al. | Yield formation strategies of a loose-panicle super hybrid rice | |
Toshkina et al. | Improved methods of original and elite seed potato growing aimed at creation of a bank of healthy potato varieties in NI Kolesnikov and DP Pavlyuk farms in Novgorod region | |
Bekusarova et al. | On the effect of PABA on germination | |
ABDOULAYE et al. | Production of potato (Solanum tuberosum) pre-basic seed through tissue culture in Katibougou, Mali | |
CN111011280A (en) | Composite culture method for shrimps and algae | |
Pomper et al. | Container production of pawpaw seedlings | |
CN107535277A (en) | A kind of breeding method of mango detoxic seedling | |
BELKHODJA | FACTOR IN RESISTANCE TO WATER STRESS IN REINA MORA VARIETY OF BEAN (Vicia faba L.) Yasmina BIDAI, Fatima Zohra MENDLI, Amina KADIRI, Asma ACHOUR | |
Bidai et al. | Organic matter as an external factor in resistance to water stress in Reina Mora variety of bean (Vicia faba L.). | |
Chauruka et al. | Management and rationalised fertilisation of improved commercial production of potato mini-tubers |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WD01 | Invention patent application deemed withdrawn after publication | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150527 |