Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (9): 142-151.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1541
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ZHAO Zhong-ying1(
), LI Qing-miao2, TIAN Meng-liang1, YANG Xiao-qian1, KANG Yao1, QIU Yu-jie1, ZHANG Qing-ling3, LIU Fan1()
Received:2020-12-22
Online:2021-09-26
Published:2021-10-25
Contact:
LIU Fan
E-mail:285461725@qq.com;liufant12006@163.com
ZHAO Zhong-ying, LI Qing-miao, TIAN Meng-liang, YANG Xiao-qian, KANG Yao, QIU Yu-jie, ZHANG Qing-ling, LIU Fan. Effects of 60Co-γ Radiation on the Seedling Rate and Plant Characteristics of Pinellia ternata Callus[J]. Biotechnology Bulletin, 2021, 37(9): 142-151.
Fig. 1 Inhibition of bud differentiation
Fig. 2 Buds differentiated normally
Fig. 3 Growth of regenerated plants treated with different radiation doses a：0 Gy treated;b：15 Gy treated;c：20 Gy treated;d：25 Gy treated;e：30 Gy treated
| 辐射剂量 Radiation dosage/Gy | 芽分化率Bud differentiation rate/% | 成苗率 Seedling rate/% |
|---|---|---|
| 0 | 100±0.00a | 100±0.00a |
| 15 | 72±6.93b | 72±6.93b |
| 20 | 63±9.02c | 63±9.02c |
| 25 | 37±4.16d | 37±4.16d |
| 30 | 30±0.00d | 30±0.00d |
Table 1 Bud differentiation rate and relative bud differen-tiation rate of Pinellia ternata callus treated with different radiation doses
| 辐射剂量 Radiation dosage/Gy | 芽分化率Bud differentiation rate/% | 成苗率 Seedling rate/% |
|---|---|---|
| 0 | 100±0.00a | 100±0.00a |
| 15 | 72±6.93b | 72±6.93b |
| 20 | 63±9.02c | 63±9.02c |
| 25 | 37±4.16d | 37±4.16d |
| 30 | 30±0.00d | 30±0.00d |
Fig. 4 High mutation frequency of regenerated plants treated with different radiation doses
Fig. 5 Variation coefficient of plant height of regenerated plants treated with different radiation doses
Fig. 6 Tuber size and fresh weight mutation frequency of regenerated plants treated with different radiation doses
Fig. 7 Variation coefficient of tuber size and fresh weight of regenerated plants treated with different radia-tion doses
Fig. 8 Leaf type mutation frequency of regenerated plants treated with different radiation doses
Fig. 9 Variation of progeny leaf shape after irradiation a：Not treated with radiation;b-f：Leaf mutation
Fig. 10 Bead mutation frequency of regenerated plants treated with different radiation doses
Fig. 11 Variation of bead buds after irradiation a：Single buds,bead buds in the middle of petiole. b：Single buds,bead buds located at the base of leaf. c：Double beadles,which are located in the middle of petiole and base of leaf
Fig. 12 Chlorophyll variation coefficient under different radiation doses
| 处理组Treatment group | 变异频率Mutation frequency/% | 变异植株数Number of mutant plants /Plant |
|---|---|---|
| 15 Gy | 10.9±7.9a | 11 |
| 20 Gy | 8.6±4.1a | 8 |
| 25 Gy | 14.3±20.8a | 9 |
| 30 Gy | 12.2±7.7a | 6 |
Table 2 Screening results of chlorophyll content under different radiation doses
| 处理组Treatment group | 变异频率Mutation frequency/% | 变异植株数Number of mutant plants /Plant |
|---|---|---|
| 15 Gy | 10.9±7.9a | 11 |
| 20 Gy | 8.6±4.1a | 8 |
| 25 Gy | 14.3±20.8a | 9 |
| 30 Gy | 12.2±7.7a | 6 |
Fig. 13 EST isozyme map a：From left to right,it is 0Gy,15Gy-4,15Gy-9,15Gy-10,15Gy-14,15Gy-17,15Gy-27,15Gy-51,15Gy-58,15Gy-74,15Gy-81,15Gy-84;b：from left to right,it is 25Gy-8,25Gy-6,20Gy-88,20Gy-73,20Gy-64,20Gy-60,20Gy-8,20Gy-7,20Gy-5,20Gy-1,0Gy;c：from left to right,it is 0Gy,25Gy-9,25Gy-13,25Gy-16,25Gy-28,25Gy-43,30Gy-1,30Gy-15,30Gy-20,30Gy-28,30Gy-32,30Gy-34
| 样品编号 Sample No. | 迁移率Mobility | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 0.09 | 0.86 | 0.91 | 0.38 | 0.9 | 0.95 | 0.47 | 0.51 | 0.53 | 0.57 | 0.56 | 0.81 | 0.6 | |
| 0Gy | + | + | — | — | — | — | — | — | — | — | — | — | — |
| 15Gy-9 | + | + | + | — | — | — | — | — | — | — | — | — | — |
| 15Gy-17 | + | + | + | + | — | — | — | — | — | — | — | — | — |
| 20Gy-1 | + | + | — | — | + | — | — | — | — | — | — | — | — |
| 20Gy-5 | + | + | — | — | + | — | — | — | — | — | — | — | — |
| 25Gy-9 | + | + | — | — | — | + | — | — | — | — | — | — | — |
| 25Gy-13 | + | + | — | — | — | + | + | + | — | — | — | — | — |
| 25Gy-16 | + | + | — | — | — | + | + | + | — | — | — | — | — |
| 25Gy-28 | + | + | — | — | — | — | — | — | + | + | — | — | — |
| 25Gy-43 | + | + | — | — | — | — | — | — | + | + | — | — | — |
| 30Gy-1 | + | + | — | — | — | + | — | — | — | — | + | — | + |
| 30Gy-28 | + | + | — | — | — | — | — | — | + | + | — | — | — |
| 30Gy-32 | + | + | — | — | — | — | — | — | — | — | — | + | — |
| 30Gy-34 | + | + | — | — | — | — | — | — | — | — | — | + | — |
Table 3 EST isozyme band number and migration rate of mutant plants
| 样品编号 Sample No. | 迁移率Mobility | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 0.09 | 0.86 | 0.91 | 0.38 | 0.9 | 0.95 | 0.47 | 0.51 | 0.53 | 0.57 | 0.56 | 0.81 | 0.6 | |
| 0Gy | + | + | — | — | — | — | — | — | — | — | — | — | — |
| 15Gy-9 | + | + | + | — | — | — | — | — | — | — | — | — | — |
| 15Gy-17 | + | + | + | + | — | — | — | — | — | — | — | — | — |
| 20Gy-1 | + | + | — | — | + | — | — | — | — | — | — | — | — |
| 20Gy-5 | + | + | — | — | + | — | — | — | — | — | — | — | — |
| 25Gy-9 | + | + | — | — | — | + | — | — | — | — | — | — | — |
| 25Gy-13 | + | + | — | — | — | + | + | + | — | — | — | — | — |
| 25Gy-16 | + | + | — | — | — | + | + | + | — | — | — | — | — |
| 25Gy-28 | + | + | — | — | — | — | — | — | + | + | — | — | — |
| 25Gy-43 | + | + | — | — | — | — | — | — | + | + | — | — | — |
| 30Gy-1 | + | + | — | — | — | + | — | — | — | — | + | — | + |
| 30Gy-28 | + | + | — | — | — | — | — | — | + | + | — | — | — |
| 30Gy-32 | + | + | — | — | — | — | — | — | — | — | — | + | — |
| 30Gy-34 | + | + | — | — | — | — | — | — | — | — | — | + | — |
| 处理组Treatment group | 变异频率Mutation frequency/% | 筛选变异植株数Screening number of mutant plants /Plants |
|---|---|---|
| 15 Gy | 2.2±3.9a | 2 |
| 20 Gy | 2.2±3.9a | 2 |
| 25 Gy | 7.9±9.9a | 5 |
| 30 Gy | 8.6±1.6a | 4 |
Table 4 Screening results of EST isozymes under different radiation treatments
| 处理组Treatment group | 变异频率Mutation frequency/% | 筛选变异植株数Screening number of mutant plants /Plants |
|---|---|---|
| 15 Gy | 2.2±3.9a | 2 |
| 20 Gy | 2.2±3.9a | 2 |
| 25 Gy | 7.9±9.9a | 5 |
| 30 Gy | 8.6±1.6a | 4 |
| 突变植株编号 Mutant plant No. | 株高 Plant height/cm | 块茎大小 Tuber size/cm | 块茎鲜重 Tuber fresh weight/g | 叶型 Leaf type | 珠芽着生位置及数目 The position and number of buds | 叶绿素含量Chlorophyll content/（mg·cm-2） | 酯酶同工酶酶带数 Number of esterase isozyme bands/Strips |
|---|---|---|---|---|---|---|---|
| 0Gy | 14-17 | 0.6-0.8 | 0.17-0.22 | 阔卵形,叶端急尖 | 仅有一个珠芽,位于半夏叶柄中部 | 0.15-0.2 | 2 |
| 15Gy-9 | 7.5 | 0.48 | 0.27 | 披针形 | 仅有一个珠芽,位于半夏叶片基部 | 0.328 | 3 |
| 15Gy-17 | 6 | 0.38 | 0.28 | 阔椭圆形 | 仅有一个珠芽,位于半夏叶片基部 | 0.351 | 4 |
| 20Gy-1 | 9.2 | 0.43 | 0.14 | 披针形 | 双珠芽,珠芽位置位于叶片基部及叶柄中部 | 0.306 | 3 |
| 20Gy-5 | 4.5 | 0.42 | 0.25 | 阔卵形,叶端骤尖 | 仅有一个珠芽,位于半夏叶片基部 | 0.327 | 3 |
| 25Gy-9 | 12 | 0.38 | 0.26 | 阔卵形 | 仅有一个珠芽,位于半夏叶片基部 | 0.346 | 3 |
| 25Gy-13 | 9.2 | 0.28 | 0.25 | 阔卵形,叶端骤尖 | 仅有一个珠芽,位于半夏叶片基部 | 0.317 | 5 |
| 25Gy-16 | 4.9 | 0.21 | 0.13 | 披针形 | 仅有一个珠芽,位于半夏叶片基部 | 0.272 | 5 |
| 25Gy-28 | 8 | 0.48 | 0.29 | 阔椭圆形 | 双珠芽,珠芽位置位于叶片基部及叶柄中部 | 0.318 | 4 |
| 25Gy-43 | 11.5 | 0.46 | 0.29 | / | 仅有一个珠芽,位于半夏叶片基部 | 0.253 | 4 |
| 30Gy-1 | 4 | / | 0.30 | 披针形 | 仅有一个珠芽,位于半夏叶片基部 | 0.345 | 5 |
| 30Gy-28 | 8.3 | / | / | 阔卵形,叶端骤尖 | 仅有一个珠芽,位于半夏叶片基部 | 0.31 | 4 |
| 30Gy-32 | 4.5 | / | 0.28 | 阔卵形 | 仅有一个珠芽,位于半夏叶片基部 | 0.396 | 3 |
| 30Gy-34 | 11.5 | / | 0.15 | 阔椭圆形 | 仅有一个珠芽,位于半夏叶片基部 | 0.341 | 3 |
Table 5 Results of different characters of mutant plants were compared with those of treatment
| 突变植株编号 Mutant plant No. | 株高 Plant height/cm | 块茎大小 Tuber size/cm | 块茎鲜重 Tuber fresh weight/g | 叶型 Leaf type | 珠芽着生位置及数目 The position and number of buds | 叶绿素含量Chlorophyll content/（mg·cm-2） | 酯酶同工酶酶带数 Number of esterase isozyme bands/Strips |
|---|---|---|---|---|---|---|---|
| 0Gy | 14-17 | 0.6-0.8 | 0.17-0.22 | 阔卵形,叶端急尖 | 仅有一个珠芽,位于半夏叶柄中部 | 0.15-0.2 | 2 |
| 15Gy-9 | 7.5 | 0.48 | 0.27 | 披针形 | 仅有一个珠芽,位于半夏叶片基部 | 0.328 | 3 |
| 15Gy-17 | 6 | 0.38 | 0.28 | 阔椭圆形 | 仅有一个珠芽,位于半夏叶片基部 | 0.351 | 4 |
| 20Gy-1 | 9.2 | 0.43 | 0.14 | 披针形 | 双珠芽,珠芽位置位于叶片基部及叶柄中部 | 0.306 | 3 |
| 20Gy-5 | 4.5 | 0.42 | 0.25 | 阔卵形,叶端骤尖 | 仅有一个珠芽,位于半夏叶片基部 | 0.327 | 3 |
| 25Gy-9 | 12 | 0.38 | 0.26 | 阔卵形 | 仅有一个珠芽,位于半夏叶片基部 | 0.346 | 3 |
| 25Gy-13 | 9.2 | 0.28 | 0.25 | 阔卵形,叶端骤尖 | 仅有一个珠芽,位于半夏叶片基部 | 0.317 | 5 |
| 25Gy-16 | 4.9 | 0.21 | 0.13 | 披针形 | 仅有一个珠芽,位于半夏叶片基部 | 0.272 | 5 |
| 25Gy-28 | 8 | 0.48 | 0.29 | 阔椭圆形 | 双珠芽,珠芽位置位于叶片基部及叶柄中部 | 0.318 | 4 |
| 25Gy-43 | 11.5 | 0.46 | 0.29 | / | 仅有一个珠芽,位于半夏叶片基部 | 0.253 | 4 |
| 30Gy-1 | 4 | / | 0.30 | 披针形 | 仅有一个珠芽,位于半夏叶片基部 | 0.345 | 5 |
| 30Gy-28 | 8.3 | / | / | 阔卵形,叶端骤尖 | 仅有一个珠芽,位于半夏叶片基部 | 0.31 | 4 |
| 30Gy-32 | 4.5 | / | 0.28 | 阔卵形 | 仅有一个珠芽,位于半夏叶片基部 | 0.396 | 3 |
| 30Gy-34 | 11.5 | / | 0.15 | 阔椭圆形 | 仅有一个珠芽,位于半夏叶片基部 | 0.341 | 3 |
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