60Co-γ辐射对半夏愈伤组织成苗及植株特性的影响

doi:10.13560/j.cnki.biotech.bull.1985.2020-1541

摘要/Abstract

摘要：

为探究不同⁶⁰Co-γ 射线对半夏愈伤组织成苗率及植株特性的影响,确定⁶⁰Co-γ辐射半夏的半致死剂量。试验采用不同剂量的⁶⁰Co-γ 射线对半夏愈伤组织进行照射,分析比较不同辐射剂量下的分化率、成苗率、死亡率及植株生长、变异情况。结果表明,在0-30 Gy范围内,芽分化率与辐射剂量成负相关;回归方程计算得到半致死剂量为22.34 Gy;辐射后的部分再生植株株高、块茎大小低于对照组再生植株,且辐射后的部分再生植株块茎鲜重、叶型、珠芽着生位置、珠芽数目、叶绿素含量、酯酶同工酶均发生不定向变异。综合形态及生理生化指标,共筛选出变异植株13株,在25 Gy处理下突变频率最高,获得变异植株5株,与半致死剂量接近,为最佳辐射剂量。

关键词: 半夏愈伤组织, ⁶⁰Co-γ射线, 半致死剂量

Abstract:

The objectives are to explore the effects of different ⁶⁰Co-γ-rays on the seedling rate and plant characteristics of Pinellia ternata callus and to determine the half lethal dose of radiation treating P. ternata. The P. ternate callus was irradiated with different doses of ⁶⁰Co-γ-ray,and then the differentiation rate,seedling rate,mortality rate,plant growth and variation were analyzed and compared. The results showed that there was a negative correlation between the bud differentiation rate and radiation dose in the range of 0-30 Gy;the half lethal dose calculated by regression equation was 22.34 Gy;the plant height and tuber size of the partially regenerated plants after irradiation were lower than those of the control group,and the fresh weight of tuber,leaf type,location of bulbils,number of bulbils,chlorophyll content and esterase isozymes of the partially regenerated plants after irradiation presented non-directional variation. According to the morphological,physiological and biochemical indexes,13 mutant plants were selected. The mutation frequency was the highest under 25 Gy treatment,and 5 mutant plants were obtained,which was close to the half lethal dose and was the optimal radiation dose.

Key words: Pinellia ternate callus, ⁶⁰Co-γ-ray, half lethal dose

赵中莹, 李青苗, 田孟良, 杨小倩, 康瑶, 邱玉洁, 张庆玲, 刘帆. ⁶⁰Co-γ辐射对半夏愈伤组织成苗及植株特性的影响[J]. 生物技术通报, 2021, 37(9): 142-151.

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.

图/表 18

图1 芽分化抑制

Fig. 1 Inhibition of bud differentiation

图2 芽正常分化

Fig. 2 Buds differentiated normally

图3 不同辐射剂量处理后再生植株生长情况 a：0 Gy处理;b：15 Gy处理;c：20 Gy处理;d：25 Gy处理;e：30 Gy处理

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

表1 不同辐射剂量处理半夏愈伤组织芽分化率和相对芽分化率

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

图4 不同辐射剂量处理再生植株株高突变频率

Fig. 4 High mutation frequency of regenerated plants treated with different radiation doses

图5 不同辐射剂量处理再生植株株高变异系数

Fig. 5 Variation coefficient of plant height of regenerated plants treated with different radiation doses

图6 不同辐射剂量处理再生植株块茎大小及鲜重突变频率

Fig. 6 Tuber size and fresh weight mutation frequency of regenerated plants treated with different radiation doses

图7 不同辐射剂量处理再生植株块茎大小及鲜重变异系数

Fig. 7 Variation coefficient of tuber size and fresh weight of regenerated plants treated with different radia-tion doses

图8 不同辐射剂量处理再生植株叶型重突变频率

Fig. 8 Leaf type mutation frequency of regenerated plants treated with different radiation doses

图9 辐照处理后代叶型变异情况 a：未经辐射处理;b-f：叶突变

Fig. 9 Variation of progeny leaf shape after irradiation a：Not treated with radiation;b-f：Leaf mutation

图10 不同辐射剂量处理再生植株珠芽突变频率

Fig. 10 Bead mutation frequency of regenerated plants treated with different radiation doses

图11 辐照处理后珠芽变异情况 a：单株芽,珠芽位于叶柄中部;b：单株芽,珠芽位于叶片基部;c：双珠芽,珠芽位于叶柄中部及叶片基部

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

图12 不同辐射剂量处理叶绿素变异系数

Fig. 12 Chlorophyll variation coefficient under different radiation doses

表2 不同辐射剂量处理叶绿素含量变化筛选结果

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

图13 EST同工酶图谱 a：由左至右依次是0Gy、15Gy-4、15Gy-9、15Gy-10、15Gy-14、15Gy-17、15Gy-27、15Gy-51、15Gy-58、15Gy-74、15Gy-81、15Gy-84;b：由左至右依次是25Gy-8、25Gy-6、20Gy-88、20Gy-73、20Gy-64、20Gy-60、20Gy-8、20Gy-7、20Gy-5、20Gy-1、0Gy;c：由左至右依次是0Gy、25Gy-9、25Gy-13、25Gy-16、25Gy-28、25Gy-43、30Gy-1、30Gy-15、30Gy-20、30Gy-28、30Gy-32、30Gy-34

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

表3 变异植株EST同工酶酶带数及迁移率结果

Table 3 EST isozyme band number and migration rate of mutant plants

样品编号 Sample No.	迁移率Mobility
样品编号 Sample No.	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	+	+	—	—	—	—	—	—	—	—	—	+	—

表4 不同辐射处理EST同工酶筛选结果

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

表5 对照处理与突变植株各变异性状测定结果

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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⁶⁰Co-γ辐射对半夏愈伤组织成苗及植株特性的影响

Effects of ⁶⁰Co-γ Radiation on the Seedling Rate and Plant Characteristics of Pinellia ternata Callus

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