Study for citrus chromosome mapping

柑橘染色体定位研究

• 批准号: 10660030
• 负责人: KITAJIMA Akira
• 金额: $ 2.56万
• 依托单位: Kochi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660030/
• 关键词: citrus; chromosome; karyotype; Giemsa stain; CMA stain; GISH; chromosome preparation method; ヘキスト染色

For making physical map of citrus chromosomes , chromosome preparation method in which vegetative tissue of mother plant use as a material and chromosomes are relatively long and useful for karyotype analysis , and identification of chromosomes should be confirmed.1. In chromosome preparation method, using young leaves of 3-5mm, 120-180min treatment of 0.3% cellurase +0.2% pectolyase solution at 37℃ and dropping suspended cells to slide-glass were useful for relatively long chromosome prepared.2 . Citrus chromosomes were classtfied to 5 types based on CMA banding patterns . That is , CMA(+) regions in terminal of both arms and a proximal (type-A) , in terminal of one arm and a proximal (type-B) , in terminal of both arms (type-C), in terminal of one arm (type-D) and no CMA(+) region (type-E) on chromosome.3. Citrus chromosome compositions depended on CMA banding pattern were 2B+8D+8E in fingered citron, 1C+8D+9E in koji, 1A+1C+8D+8E in unshiu, 2B+2C+7D+7E in 'Washington' navel orange, 2A+1B+1C+6D+8D in 'Duncan' grapefruit, 1A+1B+5C+2D+9E in 'Tosa-Buntan' , 3A+3C+3D+9E in 'Suisho-Buntan' , 2A+1B+3C+3D+9E in ' Ban-okan ' , 1A+2B+2C+4D+8E+1A or B in 'Banpeiyu ' and 4B+8D+6E in triforiate orange.4. For 'Nankan N0.20' unshiu chromosomes, genomic in situ hybridization (GISH) was performed using Dig labeled 'Nankan N0.20 ' DNA and Biotin labeled 'Tosa-Buntan ' DNA, and chromosomes were stained with anti-Dig rhodamine and avidin FITC. Eight type-D chromosomes of ' Nankan N0.20 ' were classified to I chromosome with large yellow signal, I with small yellow signal, 3 with large orange signal, I with small orange signal and 2 with small red signal. Furthermore in 'Nankan N0.20 ' chromosomes, 8 type-E chromosomes were classified to 2 long, 2 medium and 4 short chromosomes by Giemsa staining.

为制作柑桔染色体物理图谱，应确定以母本营养组织为材料，染色体较长，便于核型分析和染色体鉴定的染色体制备方法.在染色体制备方法中，以3- 5 mm的幼叶为材料，0.3%纤维素酶+0.2%果胶酶溶液在37℃下处理120- 180 min，并将悬浮细胞滴于玻片上，可获得较长的染色体.根据CMA带型将柑桔染色体分为5类。在染色体上，CMA（+）区位于两臂末端和近端（A型）、一臂末端和近端（B型）、两臂末端（C型）、一臂末端（D型）和无CMA（+）区（E型）.由CMA带型决定的柑橘染色体组成为：佛手2B+8 D +8 E，曲1C+8 D +9 E，温秀1A+1C+8 D +8 E，'华盛顿'脐橙子2B+2C+7 D +7 E，'邓肯'葡萄柚2A+1B+1C+6D+8 D，'土佐文丹' 1A+1B+5C+2D+9 E，'水照文丹' 3A+3C+3D+9 E，'半冈'为2A+1 B +3C+3D+9 E，'半培玉'为1A+2 B +2C+4D+8 E +1A或B，三福橙为4 B +8D+6 E.对于'Nankan N0.20' unshiu染色体，使用Dig标记的'Nankan N0.20 ' DNA和生物素标记的'Tosa-Buntan ' DNA进行基因组原位杂交（GISH），并用抗Dig罗丹明和亲和素FITC对染色体进行染色。南勘N0.20的8条D型染色体分别为I号黄色大信号染色体、I号黄色小信号染色体、3号橙子大信号染色体、I号橙子小信号染色体和2号红色小信号染色体。此外，在'南看N0.20 '中，8条E型染色体经Giemsa染色分为2条长染色体、2条中染色体和4条短染色体。

项目成果

Befu, M., A. Kitajima et al.: "Classification of 'Tosa-Buntan' pummelo (Citrus grandis [L.] Osb.), 'Washington' navel orange (C. sinensis [L.] Osb.) and trifoliate orange (Poncirus trifoliata [L.] Raf.) chromosomes using young leaves."J. Japan. Soc. Hort.

Befu, M., A. Kitajima 等人：“‘Tosa-Buntan’柚子 (Citrus grandis [L.] Osb.)、‘华盛顿’脐橙 (C. sinensis [L.] Osb.) 和三叶柚的分类

北島宣、別府まゆみ、日高佳子、長谷川耕二郎: "カンキツ染色体標本作製における酵素処理条件とウンシュウミカンおよびブンタン染色体の識別"園芸学会中四国支部要旨. 38. 16 (1999)

Norihiro Kitajima、Mayumi Beppu、Yoshiko Hidaka 和 Kojiro Hasekawa：“制备柑橘染色体样本的酶处理条件以及温州柑橘和 Buntan 染色体的鉴定”园艺学会中国四国分会摘要 38. 16 (1999)。

Befu,M.,Kitajima et al.: "Classification 'Tosa-Buntan' pummelo(Citrus grandis[L.]Osb.),'Washington' navel orange(C.sinensis[L.]Osb.) d trifoliate orange(Poncirus trifoliata[L.]Raf.)chromosomes using young leaves"J.Japan.Soc.Hort.Sci.. 69. 22-28 (2000)

Befu,M.,Kitajima 等人：“分类‘Tosa-Buntan’柚子（Citrus grandis[L.]Osb.）、‘华盛顿’脐橙（C.sinensis[L.]Osb.）d 枳（Poncirus）

Befu, M., A. Kitajima et al.: "Classification of 'Tosa-Bintan' purmmelo , 'Washington Navel' orange and trifoliate orange chromosomes using young leaves"J. Japan. Soc. Hort. Sci.. 67(Suppl. 2). 167-167 (1998)

Befu, M., A. Kitajima 等人：“利用幼叶对‘土佐民丹岛’红果、‘华盛顿脐橙’和三叶橙染色体进行分类”J.

Befu,M.,A.Kitajima and K.Hasegawa: "Classification of 'Tosa-Buntan' pummelo (Ctrus grandis [L.]Osb.),'Washington' Navel orange (C.sinensis [L.] Osb) and trifoliaFe orange (Poucirus tnfoliata[L.] Raf.) chroiuosoiues."J.Japan.Soc.Hort.Sci.. 69・1. 22-28 (200

Befu,M.,A.Kitajima 和 K.Hasekawa：“‘Tosa-Buntan’柚子 (Ctrus grandis [L.]Osb.)、‘华盛顿’脐橙 (C.sinensis [L.]Osb.) 和trifoliaFe 橙 (Poucirus tnfoliata[L.] Raf.) chroiuosoiues."J.Japan.Soc.Hort.Sci.. 69・1. 22-28 (200