Crops ›› 2018, Vol. 34 ›› Issue (1): 66-70.doi: 10.16035/j.issn.1001-7283.2018.01.010

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Selection Mutant of Resistant to Black Shank on Tobacco by Anther Culture

Dai Linjian1,Chen Wu1,Zhou Tian1,2,Zhong Jun1   

  1. 1 College of Agronomy, Hunan Agricultural University, Changsha 410128, Hunan, China
    2 Alleviation and Development Bureau on Liuzhi Special District of Liupanshui City of Guizhou Province, Liupanshui 553400, Guizhou, China
  • Received:2017-07-31 Revised:2017-10-16 Online:2018-02-20 Published:2018-08-24

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Abstract:

Tobacco strains GK2 were regenerated from anther culture on a selective medium containing different concentration of pathogenic active components of fermentation broth on black shank fungus. Resistance of the plants which tolerant to black shank was tested by in vitro leaf method and these plants showed different resistance levels. The disease resistance of the progenies of the plants was retested in the following two generations under mechanically inoculated and natural conditions, receptively, and all of them showed consistant, stable and heritable resistance to black shank. The agronomic characters of the resistant lines were also evaluated in the field trial in 2016-2017 and most of them averaged better than that on control GK2 plants and stably maintained. Therefore, we could conclude that we had selected resistance tobacco lines from highly potassium GK2 lines, and would be popularly applied in the field.

Key words: Flue-cured tobacco, Anther culture, Mutant, Black shank

Table 1

Difference of necrotic spot area in HR of tobacoo leaves under different treatments of fermented broth of black shank fungus cm2"

处理Treatment 最大面积Maximum area 最小面积Minimum area 平均值±标准差Mean±standard deviation
原液Stoste (C1) 0.70 0.51 0.55±0.04cB
超滤液Ultrafiltration (C2) 1.17 0.79 0.87±0.10bA
95%乙醇洗脱液Ethanol eluent (C3) 1.34 0.78 0.93±0.05aA

Table 2

Effects of different concentrations of active components of fermented broth of black shank fungus on anther germination rate of flue cured tobacco GK2 in high potassium"

浓度(%)
Concentration		 接种花药数
Inoculation anthers		 萌发出苗数
Germination		 萌发率(%)
Germination rate		 差异显著性Significance of difference
5% 1%
0(CK) 75 29 38.7 a A
1 75 22 29.3 b B
2 75 19 25.3 b B
3 75 14 18.7 b B
4 75 6 8.0 c C
5 75 5 6.7 c C

Fig.1

Chloroplast numbers in stoma guard cells of anther cultured seedlings with different ploidy|||The arrow points to the chloroplast in the stoma guard cell"

Table 3

R-1 plants disease resistance detection"

材料
Material		 株数
Plant number		 发病率(%)
Morbidity rate		 发病指数
Morbidity index		 较GK2-CK发病指数降低率(%)
Decrease rate of morbidity index compaired with GK2-CK
人工接种 GK2-D1 30 96aA 51.5bB 42.52
Artificial inoculation GK2-D2 30 90aA 48.7bB 45.65
GK2-D3 30 89aA 46.9bB 47.66
GK2-D4 30 93aA 49.3bB 44.98
GK2-D5 30 94aA 50.6bB 43.53
GK2-D6 30 92aA 49.2bB 45.09
GK2-CK 30 100aA 89.6aA -
自然发病 GK2-D1 100 79bB 31.6bB 58.31
Natural occurrence GK2-D2 100 71bB 23.5bB 68.99
GK2-D3 100 78bB 20.4bB 73.09
GK2-D4 100 75bB 28.6bB 62.27
GK2-D5 100 76bB 30.1bB 60.29
GK2-D6 100 73bB 27.9bB 63.19
GK2-CK 100 100aA 75.8aA -

Table 4

R-2 plants disease resistance detection under field conditions"

材料
Material		 发病率Morbidity rate (%) 发病指数
Morbidity index		 较GK2-CK发病指数降低率(%)
Decrease rate of morbidity
index compaired with GK2-CK
Ⅰ Ⅱ Ⅲ 平均Mean
GK2-D2 60.4 63.8 61.2 61.80bB 27.3bB 64.68
GK2-D3 66.4 64.1 63.9 64.80bB 30.8bB 60.15
GK2-D6 64.2 63.9 63.8 63.97bB 29.6bB 61.71
GK2-CK 90.6 89.7 91.4 90.57aA 77.3aA -
Coker371
(抗病对照Disease resistant control)		 56.9
 51.3
 53.7
 53.90bcB
 20.4cB
 -

Table 5

R-2 plants agronomic traits detection under field conditions (mature period)"

材料Material 株高Plant height (cm) 节距Pitch (cm) 茎围Stem girth (cm) 最大叶面积Maximum leaf area (cm2) 有效叶片数Effective blades
GK2-D2 113.40aA 26.83aA 10.92aA 2 026.40aA 19aA
GK2-D3 102.62aA 24.24aA 10.28aA 2 085.84aA 20aA
GK2-D6 112.23aA 25.23aA 11.65aA 2 012.80aA 19aA
GK2-CK 99.20aA 19.40bB 8.87bB 1 824.00bB 18aA
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