小麦—中间偃麦草（Elytrigia intermedium）双体异附加系的选育与鉴定
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摘要
中间偃麦草(Elytrigia intermedium，2n=6x=42)是禾本科小麦亚族中的多年生野生草本植物，含有许多对小麦品种改良有益的基因。创建小麦异附加系是利用中间偃麦草有益基因的重要途径。本研究综合运用形态学、细胞遗传学、生化和RAPD分子标记对已育成的10个小麦—中间偃麦草双体异附加系进行了鉴定；同时对普通小麦品种烟农15与八倍体小偃麦小偃257的杂交后代BC_1F_1、BC_1F_1~*、BC_1F_2、BC_1F_2~*和F_2进行了细胞遗传学分析，并在此基础上进行了新的双体异附加系的选育。获得主要结果如下：
1．通过细胞学和细胞遗传学分析证明，已育成的10个小麦—中间偃麦草双体异附加系具有良好的细胞学稳定性，外源染色体传递频率较高，它们的根尖细胞染色体数目为2n=44，花粉母细胞减数分裂中期Ⅰ(PMC MI)的染色体构型为2n=22Ⅱ；在主要形态特征和有关农艺性状方面10个异附加系均与亲本表现出不同程度的差异，其中Line1和Line15对白粉病表现为高抗到免疫，其抗病性来自于附加的中间偃麦草染色体。
2．通过形态性状分析、生化鉴定和测交异同性鉴定，明确了在供试的10个双体异附加系中，Line5、Line6、Line10和Line14附加的是同一对中间偃麦草染色体(可能来自第1部分同源群)；Line12和Line13附加的是另外一对中间偃麦草染色体(可能来自第2部分同源群)；Line1和Line15附加的外源染色体不同，但可能同属于第7部分同源群；Line4和Line11可能在附加外源染色体的基础上又发生了其它形式的染色体变异，对此需要进一步鉴定。
3．用200个随机引物对10个双体异附加系及其亲本进行了RAPD分析，共筛选到7个特异的RAPD引物S20、S21、S29、S57、S121、S152和S188，它们能在附加系和亲本间扩增出多态性差异谱带；利用筛选的特异引物进行扩增，建立了不同异附加系的RAPD标记，可以作为异附加系所附加外源染色体的遗传标记。
4．通过对烟农15与小偃257不同杂种世代(BC_1F_1、BC_1F_1~*、BC_1F_2、BC_1F_2~*和F_2)随机选取单株进行PMC MI染色体构型分析表明：烟农15
丁斌：小麦一中间堰麦草双休异附加系的选育与鉴定
和小堰 257分别与其FI回交产生的 BCIFI和 BCIFI”植株染色体数目在
21V＝40－46之间，单价体频率较高，并有多价体出现：90％以上的BCIFZ
和 BCIFZ“植株染色体数目为 Zpp 4144，较 BCIF1禾 BC；F厂更加趋向普通
小麦，而且单价体、多价体频率以及相对紊乱系数明显降低，同时二价体
数目有所增加。在两个回交世代中，以小堰257为回交亲本后代的平均染
色体构型要比以烟农15为回交亲本后代的平均染色体构型复杂，在单价
体数、二价体数、平均染色体数以及相对紊乱系数等方面均高于后者。
F仔 BC；F；（BC；F；、虽同属杂种第二代，但前者染色体构型要比后者复 入
杂，染色体数目变化范围较大，染色体数目在2n＝＝40－52之间，单价体数
目和相对紊乱系数都高于后者。
5．通过减数分裂染色体构型分析，在烟农15与小僵257不同杂种世
代（BCIFI、BCIFI、BCIFZ、BCIFZ禾 FZ）共选育出 16个附加不同数目
外源染色体的植株。其中单体异附加植株（Zn＝21 11＋I）和双单体异附
加植株Q 11＋I＋I）各 4个，其它类型单体异附加植株 6个；选
育出双体异附加植株（2n＝＝22 11）二个。不同类型附加株在各世代出现频
率差异较大，其中双体异附加植株全部出现在BCIFZ，说明BCIFZ与其它
世代相比更有利于双体异附加系的选择。
Elytrigia intermedium carries a large number of valuable genes for wheat improvement. It is an important way of using these beneficial genes by creating alien addition lines. In this study, 10 alien disomic addition lines obtained from common wheat v-Elytrigia intermedium were identified with morphological markers, cytological markers, biochemical markers and RAPD markers. Wheat-Elytrigia intermedium alien addition lines were isolated from five different cross generations from common wheat Yannongl5 x Xiaoyan257 on the basis of cytogenetic analysis. The results are as follows:
1. 10 alien disomic additon lines were stable in heredity. The number of chromosomes of root tip cells was 44, and the chromosome configuration at PMC M I was 2n=22llin these alien disomic addition lines. Main morphological characteristics were different between 10 alien disomic addition lines and their parents. Among them, linel and line 15 could resist powdery mildew effectively, and the resistance came from additive chromosomes of Elytrigia intermedium.
2. Synthetic analysis by using morphological characteristics, biochemical identification and half diallel cross indicated that Iine5, Iine6, linelO and Iinel4 might contain Elytrigia intermedium chromosomes of homoeologous group 1; The added alien chromosomes of Linel2 and Iinel3 might belong to homoeologous group 2; Linel and line 15 might be different in additive chromosomes which might belong to homoeologous group 7; Other chromosome aberration might also occur in Line4 and linel 1, and this needs further study.
3. 200 random primers were used for RAPD amplification, the specific bands of Elytrigia intermedium parent could be observed in the electrophoretogram of 10 alien disomic addition lines by 7 of them (S20, S21, S29, S57, S121, SI52 and SI88), and RAPD markers for alien addition chromosomes were set up.
4. The chromosome configurations of 5 different generations from Yannongl5 X Xiaoyan257 were analyzed. The results showed that the range of chromosome numbers in BCiF| and BCiFi* was 40-46, and the frequencies of univalents were high. More than 90% individual plants of BC1F2 and BC1F2' carried 41-44 chromosomes, which more tended to common wheat than BC1F1 and BC1F1. Moreover, the frequencies of
univalents, multivalents and RCC declined significantly in them, whose bivalents added at the same time. The mean chromosome configurations of individual plants when backcross parent was Xiaoyan257 were more complex in the number of univalents, bivalents, average chromosomes and RCC than when backcross parent was YannonglS. F2 had more complex chromosome configurations with wide rage of chromosomes (40-52) and higher number of univalents and RCC than BC1F1 (BC1F1*).
5. Based on cytogenetic analysis, 16 individual plants with different number of alien chromosomes were obtained from different cross generations (BC1F1, BC1F1 , BC1F2, BC1F2* and F2). Among them, the number of plants with chromosome configurations of 2n=21II+ I and 2n=21II+ I + I was 4 respectively, and the number of other monosomic addition types was 6. At the same time, 2 plants with chromosome configuration of 2n=22 II were obtained in BCi12. The frequencies of different alien addition plants were different in the several cross generations, and BQF2 was more efficient for the selecting of alien disomic addition lines.
1．通过细胞学和细胞遗传学分析证明，已育成的10个小麦—中间偃麦草双体异附加系具有良好的细胞学稳定性，外源染色体传递频率较高，它们的根尖细胞染色体数目为2n=44，花粉母细胞减数分裂中期Ⅰ(PMC MI)的染色体构型为2n=22Ⅱ；在主要形态特征和有关农艺性状方面10个异附加系均与亲本表现出不同程度的差异，其中Line1和Line15对白粉病表现为高抗到免疫，其抗病性来自于附加的中间偃麦草染色体。
2．通过形态性状分析、生化鉴定和测交异同性鉴定，明确了在供试的10个双体异附加系中，Line5、Line6、Line10和Line14附加的是同一对中间偃麦草染色体(可能来自第1部分同源群)；Line12和Line13附加的是另外一对中间偃麦草染色体(可能来自第2部分同源群)；Line1和Line15附加的外源染色体不同，但可能同属于第7部分同源群；Line4和Line11可能在附加外源染色体的基础上又发生了其它形式的染色体变异，对此需要进一步鉴定。
3．用200个随机引物对10个双体异附加系及其亲本进行了RAPD分析，共筛选到7个特异的RAPD引物S20、S21、S29、S57、S121、S152和S188，它们能在附加系和亲本间扩增出多态性差异谱带；利用筛选的特异引物进行扩增，建立了不同异附加系的RAPD标记，可以作为异附加系所附加外源染色体的遗传标记。
4．通过对烟农15与小偃257不同杂种世代(BC_1F_1、BC_1F_1~*、BC_1F_2、BC_1F_2~*和F_2)随机选取单株进行PMC MI染色体构型分析表明：烟农15
丁斌：小麦一中间堰麦草双休异附加系的选育与鉴定
和小堰 257分别与其FI回交产生的 BCIFI和 BCIFI”植株染色体数目在
21V＝40－46之间，单价体频率较高，并有多价体出现：90％以上的BCIFZ
和 BCIFZ“植株染色体数目为 Zpp 4144，较 BCIF1禾 BC；F厂更加趋向普通
小麦，而且单价体、多价体频率以及相对紊乱系数明显降低，同时二价体
数目有所增加。在两个回交世代中，以小堰257为回交亲本后代的平均染
色体构型要比以烟农15为回交亲本后代的平均染色体构型复杂，在单价
体数、二价体数、平均染色体数以及相对紊乱系数等方面均高于后者。
F仔 BC；F；（BC；F；、虽同属杂种第二代，但前者染色体构型要比后者复 入
杂，染色体数目变化范围较大，染色体数目在2n＝＝40－52之间，单价体数
目和相对紊乱系数都高于后者。
5．通过减数分裂染色体构型分析，在烟农15与小僵257不同杂种世
代（BCIFI、BCIFI、BCIFZ、BCIFZ禾 FZ）共选育出 16个附加不同数目
外源染色体的植株。其中单体异附加植株（Zn＝21 11＋I）和双单体异附
加植株Q 11＋I＋I）各 4个，其它类型单体异附加植株 6个；选
育出双体异附加植株（2n＝＝22 11）二个。不同类型附加株在各世代出现频
率差异较大，其中双体异附加植株全部出现在BCIFZ，说明BCIFZ与其它
世代相比更有利于双体异附加系的选择。
Elytrigia intermedium carries a large number of valuable genes for wheat improvement. It is an important way of using these beneficial genes by creating alien addition lines. In this study, 10 alien disomic addition lines obtained from common wheat v-Elytrigia intermedium were identified with morphological markers, cytological markers, biochemical markers and RAPD markers. Wheat-Elytrigia intermedium alien addition lines were isolated from five different cross generations from common wheat Yannongl5 x Xiaoyan257 on the basis of cytogenetic analysis. The results are as follows:
1. 10 alien disomic additon lines were stable in heredity. The number of chromosomes of root tip cells was 44, and the chromosome configuration at PMC M I was 2n=22llin these alien disomic addition lines. Main morphological characteristics were different between 10 alien disomic addition lines and their parents. Among them, linel and line 15 could resist powdery mildew effectively, and the resistance came from additive chromosomes of Elytrigia intermedium.
2. Synthetic analysis by using morphological characteristics, biochemical identification and half diallel cross indicated that Iine5, Iine6, linelO and Iinel4 might contain Elytrigia intermedium chromosomes of homoeologous group 1; The added alien chromosomes of Linel2 and Iinel3 might belong to homoeologous group 2; Linel and line 15 might be different in additive chromosomes which might belong to homoeologous group 7; Other chromosome aberration might also occur in Line4 and linel 1, and this needs further study.
3. 200 random primers were used for RAPD amplification, the specific bands of Elytrigia intermedium parent could be observed in the electrophoretogram of 10 alien disomic addition lines by 7 of them (S20, S21, S29, S57, S121, SI52 and SI88), and RAPD markers for alien addition chromosomes were set up.
4. The chromosome configurations of 5 different generations from Yannongl5 X Xiaoyan257 were analyzed. The results showed that the range of chromosome numbers in BCiF| and BCiFi* was 40-46, and the frequencies of univalents were high. More than 90% individual plants of BC1F2 and BC1F2' carried 41-44 chromosomes, which more tended to common wheat than BC1F1 and BC1F1. Moreover, the frequencies of
univalents, multivalents and RCC declined significantly in them, whose bivalents added at the same time. The mean chromosome configurations of individual plants when backcross parent was Xiaoyan257 were more complex in the number of univalents, bivalents, average chromosomes and RCC than when backcross parent was YannonglS. F2 had more complex chromosome configurations with wide rage of chromosomes (40-52) and higher number of univalents and RCC than BC1F1 (BC1F1*).
5. Based on cytogenetic analysis, 16 individual plants with different number of alien chromosomes were obtained from different cross generations (BC1F1, BC1F1 , BC1F2, BC1F2* and F2). Among them, the number of plants with chromosome configurations of 2n=21II+ I and 2n=21II+ I + I was 4 respectively, and the number of other monosomic addition types was 6. At the same time, 2 plants with chromosome configuration of 2n=22 II were obtained in BCi12. The frequencies of different alien addition plants were different in the several cross generations, and BQF2 was more efficient for the selecting of alien disomic addition lines.
引文
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