CN112931184A - 一种抗小象甲甘薯品种的鉴定方法 - Google Patents
一种抗小象甲甘薯品种的鉴定方法 Download PDFInfo
- Publication number
- CN112931184A CN112931184A CN202110309090.2A CN202110309090A CN112931184A CN 112931184 A CN112931184 A CN 112931184A CN 202110309090 A CN202110309090 A CN 202110309090A CN 112931184 A CN112931184 A CN 112931184A
- Authority
- CN
- China
- Prior art keywords
- sweet potato
- weevil
- insect
- leaves
- weevils
- 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
- 244000017020 Ipomoea batatas Species 0.000 title claims abstract description 50
- 235000002678 Ipomoea batatas Nutrition 0.000 title claims abstract description 47
- 238000000034 method Methods 0.000 title claims abstract description 32
- 241000254171 Curculionidae Species 0.000 title claims abstract description 14
- 241001503766 Cylas formicarius Species 0.000 claims abstract description 47
- 241000238631 Hexapoda Species 0.000 claims abstract description 27
- 230000009105 vegetative growth Effects 0.000 claims abstract description 8
- 235000003642 hunger Nutrition 0.000 claims description 5
- 230000037351 starvation Effects 0.000 claims description 4
- 241000196324 Embryophyta Species 0.000 abstract description 14
- 208000006877 Insect Bites and Stings Diseases 0.000 abstract description 11
- 238000011156 evaluation Methods 0.000 abstract description 7
- 239000003147 molecular marker Substances 0.000 abstract description 6
- 241000406668 Loxodonta cyclotis Species 0.000 abstract description 4
- 238000009395 breeding Methods 0.000 abstract description 4
- 230000005540 biological transmission Effects 0.000 abstract description 3
- 230000001488 breeding effect Effects 0.000 abstract description 3
- 230000020595 eating behavior Effects 0.000 abstract description 3
- 230000002068 genetic effect Effects 0.000 abstract description 3
- 238000010792 warming Methods 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 description 6
- 230000002265 prevention Effects 0.000 description 6
- 244000061456 Solanum tuberosum Species 0.000 description 5
- 235000002595 Solanum tuberosum Nutrition 0.000 description 5
- 241001012098 Omiodes indicata Species 0.000 description 4
- 241000607479 Yersinia pestis Species 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 230000006399 behavior Effects 0.000 description 3
- 241001155998 Anthonomus rubi Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 235000021186 dishes Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 241000254173 Coleoptera Species 0.000 description 1
- 241000207782 Convolvulaceae Species 0.000 description 1
- 235000021506 Ipomoea Nutrition 0.000 description 1
- 241000207783 Ipomoea Species 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 230000003031 feeding effect Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000002420 orchard Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- BULVZWIRKLYCBC-UHFFFAOYSA-N phorate Chemical compound CCOP(=S)(OCC)SCSCC BULVZWIRKLYCBC-UHFFFAOYSA-N 0.000 description 1
- 235000012015 potatoes Nutrition 0.000 description 1
- 230000008261 resistance mechanism Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Botany (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
本发明公开了一种抗小象甲甘薯品种的鉴定方法。在甘薯营养生长期的叶片上接待测甘薯小象甲,让甘薯小象甲取食,然后将甘薯小象甲剥离叶片,统计虫食面积,根据虫咬面积大小判断甘薯小象甲是否为抗小象甲甘薯品种。本发明利用ImageJ或其他电子图形统计软件计算灰度的方法统计虫洞透光的面积,这种方法避免了主观将小象甲为害程度分级的抗性评价方式,从而客观而直接地反应甘薯小象甲的取食行为。虫咬面积大,则植株不抗虫,虫咬面积小或不咬则表现为抗虫。量化的植株叶片虫咬面积不用进行分级,可直接作为表型数据结合遗传图谱等进行甘薯小象甲抗性QTLs定位和GWAS分析,从而促进常规育种和分子标记辅助选择甘薯小象甲抗性品种的进程。
Description
技术领域:
本发明属于农业害虫防治技术领域,具体涉及一种抗小象甲甘薯品种的鉴定方法。
背景技术:
甘薯[Ipomoea batatas(L.)Lam.]属于旋花科番薯属,是重要的粮食作物。甘薯具有粮食、蔬菜、药材、饲料和能源作物等多种用途。甘薯在世界范围内广泛栽培,其中亚洲和非洲的热带和亚热带地区是甘薯的主要种植地区,而中国和一些非洲国家是世界上主要的甘薯种植国。中国是世界上甘薯种植面积最大的国家,占全球甘薯产量的70%,而广东省是我国甘薯的重要生产区域,其种植面积约为15万公顷。得益于广东温暖的气候,广东省甘薯生产占领了全国冬春薯市场,具有可观的经济效益。然而广东高温高湿的气候环境,也导致了甘薯病虫害频繁发生,严重威胁着甘薯的种植。
甘薯小象甲(Cylas formicarius)属于鞘翅目蚁象虫科,是危害甘薯生产最严重的虫害。甘薯小象甲在热带和亚热带气候温暖的地区(广东、福建、海南、台湾等)一年可发生多代。甘薯小象甲成虫不仅危害甘薯地上部分(茎和叶),导致植物逐渐萎蔫死亡,并且在结薯期将卵产在薯块中,幼虫在薯块中蛀食,造成薯块无法储藏和食用。一般情况下,甘薯小象甲为害甘薯生产造成10%至30%的损失,严重时可导致绝收。
甘薯小象甲是备受全球关注的甘薯虫害,但长期以来抗虫品种的缺乏制约着甘薯的种植和生产。目前甘薯小象甲的防治措施主要采用非疫区检疫,农业防治、化学防治和生物防治相结合的综合防治办法,但仍以化学防治为主。据调查,在广东省湛江的甘薯种植区,农民大量施用甲拌磷等高毒农药,不仅加重了生产成本,同时存在严重的生态环境和食品安全等方面的问题。更重要的是,近年来由于种苗调运和商品薯带虫的原因,导致甘薯小象甲向长江中下游地区扩散,威胁长江中下游地区甘薯种植。因此,建立客观可量化的抗性评价方法对鉴定甘薯小象甲抗性品种具有重要意义,同时结合分子标记辅助育种的方法,解析其抗性机制,对甘薯抗小象甲品种的培育具有重要意义。此外也有利于经济适用、环境友好型甘薯种植产业的发展。
目前,抗甘薯小象甲品种评价方法多以薯块抗性评价为主,但却忽略了甘薯小象甲首先取食为害营养生长时期甘薯植株的茎和叶片,造成植株失水萎蔫枯死,无法结薯的重要事实。同时,现有的甘薯小象甲抗性评价方法多为主观定义的危害指数,人为将危害程度分级,无法利用直接客观可量化的统计方法确定为害程度,从而造成甘薯小象甲抗性表型数据不稳定,影响抗性分子标记的定位。
发明内容:
本发明的目的是提供一种抗小象甲甘薯品种的鉴定方法,为甘薯小象甲抗性品种的筛选提供一个新的思路和鉴定方法。同时客观可靠的表型鉴定方法更有利于甘薯抗小象甲分子标记的获得,从而加快分子标记辅助选择甘薯小象甲抗性品种的育种进程。
本发明的抗小象甲甘薯品种的鉴定方法,包括以下步骤:
在甘薯营养生长期的叶片上接待测甘薯小象甲,让甘薯小象甲取食,然后将甘薯小象甲剥离叶片,统计虫食面积,根据虫咬面积大小判断甘薯小象甲是否为抗小象甲甘薯品种。
一般虫咬面积大,则植株不抗虫,虫咬面积小或不咬则表现为抗虫。
优选,所述的统计虫食面积是将咬食后的叶片放在X光片观片机上拍照,利用ImageJ软件或其他电子图形统计虫食面积。
所述的甘薯营养生长期的叶片可以是离体叶片或非离体叶片。
所述的在甘薯营养生长期的叶片上接待测甘薯小象甲优选是将一片叶片放置于培养皿中,每个培养皿中放入10只甘薯小象甲成虫,置于28℃条件下取食4.5小时。
进一步优选,所述的甘薯小象甲成虫是饥饿处理24h后再接到叶片上。
优选,所述的叶片保留8mm的叶柄长度,用湿润1cm2的纸巾包住叶柄处,防止叶片失水萎蔫,同时防止小象甲取食叶柄处。
本发明利用ImageJ或其他电子图形统计软件计算灰度的方法统计虫洞透光的面积,这种方法避免了主观将小象甲为害程度分级的抗性评价方式,从而客观而直接地反应甘薯小象甲的取食行为。虫咬面积大,则植株不抗虫,虫咬面积小或不咬则表现为抗虫。量化的植株叶片虫咬面积不用进行分级,可直接作为表型数据结合遗传图谱等进行甘薯小象甲抗性QTLs定位和GWAS分析,从而促进常规育种和分子标记辅助选择甘薯小象甲抗性品种的进程。
利用本方法分析两次甘薯抗感品种杂交F1子代的叶片抗性评价实验结果发现,两次不同时期不同环境中种植的F1子代群体的抗性表型数据具有高度的相关性,表明本发明中建立的叶片甘薯小象甲抗性评价方法客观而可靠。
附图说明:
图1是本发明的鉴定方法的流程示意图。
具体实施方式:
下述实施例中所使用的试验方法,如无特殊说明均为常规试验方法。
下面将以实施例的方式对本发明作进一步的详细描述,但以下详细描述不应以限制性的意义来理解,本发明的范围仅由所附权利要求界定。
实施例1:
本实施例的抗甘薯小象甲品种的鉴定方法,包含以下操作步骤(图1):
(1)选择同一批培养的甘薯小象甲成虫放入培养皿中,每皿10只虫;
(2)提前将培养皿中的小象甲饥饿处理24h;
(3)剪取营养生长期(30天)的甘薯叶片,三个重复,一般为第3片叶至第5片叶;
(4)剪取的叶片保留8mm的叶柄长度,用湿润1cm2的纸巾包住叶柄处,防止叶片失水萎蔫,同时防止小象甲取食叶柄处;
(5)将步骤4中的叶片放入步骤2中的培养皿中,每个培养皿一片叶片,供小象甲取食;
(6)将步骤5中的培养皿放入纸箱中,置于28℃条件下饲喂4.5h;
(7)4.5h后取出培养皿,将甘薯小象甲剥离叶片,随后将咬食后的叶片放在X光片观片机上拍照,便于统计虫咬情况;
(8)利用ImageJ软件统计虫食面积,计算三个叶片重复的平均值和标准差,从而确定虫咬面积,获得甘薯小象甲抗性数据。
技术要点
步骤3中,每个甘薯植株品种培养在同一生长环境中,剪取叶片时每株甘薯植株选择同一叶位的叶片,避免了因叶片生长状况不同造成的取食行为差异。
步骤1中,由于甘薯小象甲需要提前饥饿处理24h,所以需要大量培养虫源,从而避免反复饥饿处理虫子导致虫子取食活力下降的情况。
步骤6中,通过观察小象甲取食叶片的行为发现,0-4.5h小象甲取食叶片的行为逐渐活跃,4.5h后甘薯小象甲开始停止取食行为,因此确定4.5h为统计叶片虫食面积的最佳时间点。
步骤7中,甘薯小象甲取食叶片产生的虫洞虽小,但利用X光片观片机拍照,通过调节亮度使得叶片上的虫洞能够更好地透过光。统计透光面积即能真实地反应虫咬情况。
利用上述方法对抗(N73)、感(G87)品种杂交创建的F1子代进行甘薯小象甲抗性分析,分析所用材料种植在广东省广州市中国科学院华南植物园分子生物分析及遗传改良中心试验田,于2019年11月进行抗性评价实验。如表1所示,以敏感品种‘广薯87’和抗性品种N73的叶片虫食面积作为对照,F1群体中编号为102的子代,其叶片虫食面积为16.62mm2,占叶片总面积的0.74%,其抗性数据趋向于敏感亲本‘广薯87’,故判定为敏感;F1群体中编号为19的子代,其叶片虫食面积为7.44mm2,占叶片总面积的0.31%,抗性数据趋向于抗性亲本N73,故判定为抗。
表1
品种及子代 | 虫食面积(平均值) | 虫食面积占比(平均值) | 抗性 |
广薯87 | 15.23mm<sup>2</sup> | 0.47% | 敏感 |
N73 | 8.31mm<sup>2</sup> | 0.26% | 抗 |
F<sub>1</sub>-102 | 16.62mm<sup>2</sup> | 0.74% | 敏感 |
F<sub>1</sub>-19 | 7.44mm<sup>2</sup> | 0.31% | 抗 |
Claims (6)
1.一种抗小象甲甘薯品种的鉴定方法,其特征在于,包括以下步骤:
在甘薯营养生长期的叶片上接待测甘薯小象甲,让甘薯小象甲取食,然后将甘薯小象甲剥离叶片,统计虫食面积,根据虫咬面积大小判断甘薯小象甲是否为抗小象甲甘薯品种。
2.根据权利要求1所述的鉴定方法,其特征在于,所述的统计虫食面积是将咬食后的叶片放在X光片观片机上拍照,利用ImageJ软件或其他电子图形统计虫食面积。
3.根据权利要求1所述的鉴定方法,其特征在于,所述的甘薯营养生长期的叶片是离体叶片或非离体叶片。
4.根据权利要求1、2或3所述的鉴定方法,其特征在于,所述的在甘薯营养生长期的叶片上接待测甘薯小象甲是将一片叶片放置于培养皿中,每个培养皿中放入10只甘薯小象甲成虫,置于28℃条件下取食4.5小时。
5.根据权利要求1、2或3所述的鉴定方法,其特征在于,所述的甘薯小象甲成虫是饥饿处理24h后再接到叶片上。
6.根据权利要求1、2或3所述的鉴定方法,其特征在于,所述的叶片保留8mm的叶柄长度,用湿润1cm2的纸巾包住叶柄处。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110309090.2A CN112931184A (zh) | 2021-03-23 | 2021-03-23 | 一种抗小象甲甘薯品种的鉴定方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110309090.2A CN112931184A (zh) | 2021-03-23 | 2021-03-23 | 一种抗小象甲甘薯品种的鉴定方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN112931184A true CN112931184A (zh) | 2021-06-11 |
Family
ID=76227963
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110309090.2A Pending CN112931184A (zh) | 2021-03-23 | 2021-03-23 | 一种抗小象甲甘薯品种的鉴定方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112931184A (zh) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103555813A (zh) * | 2013-09-26 | 2014-02-05 | 广东省农业科学院作物研究所 | 一种快速鉴定甘薯茎腐病抗性的方法 |
CN105191679A (zh) * | 2015-06-19 | 2015-12-30 | 广西壮族自治区农业科学院玉米研究所 | 一种抗小象甲甘薯的鉴定方法 |
-
2021
- 2021-03-23 CN CN202110309090.2A patent/CN112931184A/zh active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103555813A (zh) * | 2013-09-26 | 2014-02-05 | 广东省农业科学院作物研究所 | 一种快速鉴定甘薯茎腐病抗性的方法 |
CN105191679A (zh) * | 2015-06-19 | 2015-12-30 | 广西壮族自治区农业科学院玉米研究所 | 一种抗小象甲甘薯的鉴定方法 |
Non-Patent Citations (4)
Title |
---|
南京农业大学: "《昆虫生理生化实验》", 31 May 1993, 农业出版社 * |
官春云: "《植物育种理论与方法》", 31 January 2004 * |
滑金锋等: "小象甲抗性鉴定方法及不同品种甘薯抗性研究", 《西南农业学报》 * |
饶顺发等: "小象甲取食处理下甘薯‘广薯87’的转录组分析", 《植物生理学报》 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Barrett | Crop mimicry in weeds | |
Ceballos et al. | Adaptation of cassava to changing climates | |
Krishnan et al. | A review on Paracoccus marginatus Williams, papaya mealybug (Hemiptera: Pseudococcidae) | |
Paul et al. | Screening of maize genotypes against fall army worm, Spodoptera frugiperda (Smith) with reference to plant morphological characters at Raipur (Chhattisgarh) | |
Atkins et al. | Reaction of small-grain varieties to green bug attack | |
Mbasani-Mansi et al. | Resistance of Moroccan lentil genotypes to Orobanche crenata infestation | |
Chambers et al. | Susceptibility of hazelnut cultivars to filbertworm, Cydia latiferreana | |
Mamun et al. | Assessment of several mustard varieties resistance to mustard aphid, Lipaphis erysimi (Kalt.) | |
CN115152704A (zh) | 一种天敌释放方法及其在防治害虫方面的应用与玉米大田害虫防治方法 | |
CN112931184A (zh) | 一种抗小象甲甘薯品种的鉴定方法 | |
Ochieng et al. | A Survey of farmers’ perceptions and management strategies of the sweet potato weevil in Homa Bay County, Kenya | |
Dhillon et al. | Influence of cytoplasmic male sterility on expression of different mechanisms of resistance in sorghum to Atherigona soccata (Diptera: Muscidae) | |
Supartha et al. | Monitoring of damage and distribution of invasive fall armyworm, Spodoptera frugiperda (JE Smith)(Lepidoptera: Noctuidae) on maize crop in Karo, North Sumatera, Indonesia | |
Nasrin et al. | Evaluation of tomato varieties against resistance to fruit borer (Helicoverpa armigera Hub.). | |
EP et al. | Mendelian Inheritance in Wheat and Barley Crosses: With Probable Error Studies on Class Frequencies | |
Manjy | Estimation the population density of leafminer Liriomyza trifolii (Burgess)(Diptera: Agromyzidae) and effect the infestation and severity of infestation on some the varieties of cowpea. | |
İlçin | Investigation of Orthoptera: Insecta fauna of useful, harmful and predator species in the batman region (turkey) | |
Kabir et al. | Effects of varying temperature on the reproduction and damage potential of Heterodera schachtii to Chinese cabbage (Brassica rapa pekinensis) | |
Siregar et al. | Inventory insects of sorghum plantation in Northern Sumatera, Indonesia | |
Silva et al. | Nitrogen doses and weed control via intercropping with gliricidia for corn production | |
AbaDura | Review paper on diversity, management practices and genetic erosion of enset (E. Ventricosum (Welw) Chessman) in South and Southwest Ethiopia | |
Abdel-Aaal | A comparative Study on Some Maize Cultivars and Their Relationship to Damage Caused by Hooded Crow, Corvus corone sardonius at Qena Governorate, EGYPT. | |
OYEWALE | INCIDENCE AND SEVERITY OF PINK STALKBORER (SESAMIA CALAMISTIS HAMPSON) AND SCREENING OF MAIZE GENOTYPES FOR RESISTANCE IN SOUTHERN GUINEA SAVANNA OF NIGERIA | |
Eshetu et al. | Grain Yield Performance of Pearl Millet (Pennisetum glaucum) Genotypes in Kebribeya District of Somali Region, Ethiopia. | |
Khammar et al. | Investigating the seed and oil quantitive reactions of sunflower cultivars in different dates at Sistan region |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210611 |