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.

制作柑橘染色体物理图谱时，应采用以母株植物组织为材料的染色体制备方法，染色体较长，有利于核型分析，并确认染色体的鉴定。 1．染色体制备方法中，采用幼体。 3-5毫米的叶子，0.3％纤维素酶+0.2％果胶酶溶液在37℃下处理120-180分钟，并将悬浮细胞滴到载玻片上，可用于2．根据CMA带型将柑橘染色体分为5种类型，即双臂末端和近端的CMA(+)区、单臂末端和近端的CMA(+)区。 (B型)、双臂末端(C型)、单臂末端(D型)且染色体上无CMA(+)区域(E型)。3．取决于CMA条带图案的成分是：佛手中的2B+8D+8E，曲中的1C+8D+9E，温州中的1A+1C+8D+8E，‘华盛顿’脐橙中的2B+2C+7D+7E，2A+1B +1C+6D+8D“邓肯”葡萄柚， “土佐文谈”中的 1A+1B+5C+2D+9E、“水翔文谈”中的 3A+3C+3D+9E、“番冈”中的 2A+1B+3C+3D+9E、1A+2B+ 2C 'Banpeiyu' 中的 +4D+8E+1A 或 B 以及三叶草中的 4B+8D+6E 4. 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)，并用抗染色'南坎N0.20'的8条D型染色体经地高辛罗丹明和亲和素FITC分类为黄色大信号的I染色体、黄色小信号的I染色体、黄色大信号的3条染色体。橙色信号，I 带有小橙色信号，2 条带有小红色信号。此外，在'Nankan N0.20'染色体中，8 条E 型染色体通过吉姆萨染色被分为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.

別府まゆみ・北島宣 他: "'土佐文旦','ワシントンネーブル'オレンジおよびカラタチの幼葉を用いた染色体の識別" 園芸学会雑誌. 67(別2). 167 (1998)

Mayumi Beppu、Nobuaki Kitajima 等人：“利用‘土佐柚子’、‘华盛顿脐橙’和日本三叶草的嫩叶鉴定染色体”，日本园艺学会杂志 67（第 2 部分）。