南京林业大学学报(自然科学版) ›› 2020, Vol. 44 ›› Issue (1): 54-62.doi: 10.3969/j.issn.1000-2006.201904002
荆茹月1(), 王佩兰1, 黄振2, 何林骏1, 李志辉1,*()
收稿日期:
2019-04-01
修回日期:
2019-06-20
出版日期:
2020-02-08
发布日期:
2020-02-02
通讯作者:
李志辉
作者简介:
荆茹月( 基金资助:
JING Ruyue1(), WANG Peilan1, HUANG Zhen2, HE Linjun1, LI Zhihui1,*()
Received:
2019-04-01
Revised:
2019-06-20
Online:
2020-02-08
Published:
2020-02-02
Contact:
LI Zhihui
摘要:
【目的】 探究渗透调节物质种类及其不同浓度对香樟体细胞胚胎发生及植株再生的影响,为提高香樟体胚诱导率、优化体细胞胚胎发生体系奠定理论基础。【方法】 以7月初在湖南省郴州、长沙、娄底及怀化市采集的香樟(Cinnamomum camphora L.)的未成熟合子胚为试验材料,分析不同碳源种类、浓度及组合方式对香樟体胚诱导、芽萌发、生根等方面的影响。【结果】 适合香樟的体胚诱导培养基为MS+1.0 mg/L的6-苄氨基腺嘌呤(6-BA)+0.3 mg/L的2,4-二氯苯氧乙酸(2,4-D)+1.0 mg/L 链霉素+15 g/L 蔗糖+30 g/L 山梨醇+6.5 g/L 琼脂+700 mg/L水解酪蛋白;增殖培养基为MS+15 g/L 蔗糖+30 g/L 山梨醇+6.5 g/L 琼脂+700 mg/L 水解酪蛋白;芽萌发培养基为MS+0.1 mg/L 苯基噻二唑基脲(TDZ)+0.5 mg/L 萘乙酸(NAA)+0.1 mg/L 吲哚丁酸(IBA)+15 g/L 蔗糖+15 g/L 山梨醇+6.5 g/L 琼脂;生根培养基为MS+1.0 mg/L IBA+15 g/L 蔗糖+15 g/L 山梨醇+6.5 g/L 琼脂。【结论】 经过改良,添加15 g/L蔗糖和30 g/L山梨醇的组合碳源,香樟体胚的诱导率明显提高;添加15 g/L蔗糖和15 g/L山梨醇的组合碳源处理的新植株生根条数多、主根明显,生长状况更优。
中图分类号:
荆茹月,王佩兰,黄振,等. 碳源对不同产地香樟体细胞胚胎发生及植株再生的影响[J]. 南京林业大学学报(自然科学版), 2020, 44(1): 54-62.
JING Ruyue, WANG Peilan, HUANG Zhen, HE Linjun, LI Zhihui. Effects of different carbon sources on somatic embryo of Cinnamomum camphora L.[J].Journal of Nanjing Forestry University (Natural Science Edition), 2020, 44(1): 54-62.DOI: 10.3969/j.issn.1000-2006.201904002.
图1
香樟体细胞胚胎发育过程的组织学观察 A-C.直接发生方式产生的再生植株regenerated plants produced by direct occurrence. A. 子叶胚cotyledon embryos; B. 子叶胚发芽germination of cotyledonary embryos; C. 子叶胚生根长成新植株cotyledon embryos grow into new plants. D-I. 间接发生方式产生的再生植株regenerated plants produced by indirect occurrence. D. 愈伤组织callus; E. 胚性愈伤组织embryonic callus; F. 球形胚、心形胚(红色箭头所指为心形胚)globular embryo heart-shaped embryo (red arrow represents heart-shaped embryo); G. 成熟子叶胚上的绿色萌发点green germination point on mature cotyledon embryos; H. 成熟子叶胚生根rooting of cotyledonary embryos; I. 成熟子叶胚长出多个新植株cotyledon embryos grow into multiple new plants."
表1
不同碳源与材料产地对香樟体胚诱导的影响"
碳源种类 types of carbon sources | 体胚诱导率/% induction rate of somatic embryo | |||
---|---|---|---|---|
郴州 Chenzhou | 长沙 Changsha | 怀化 Huaihua | 娄底 Loudi | |
蔗糖sucrose | 72.9±2.6 aC | 83.7±1.1 aA | 77.4±1.5 aB | 69.1±1.3 aD |
葡萄糖glucose | 53.2±2.7 bB | 59.7±1.9 bA | 45.2±2.1 bC | 44.9±1.6 bC |
麦芽糖maltose | 0.0 | 10.3±3.9 eA | 3.5±1.7 dB | 0.0 |
山梨醇sorbitol | 33.3±1.5 cC | 45.3±1.3 cA | 46.4±1.9 bA | 37.9±1.1 cB |
甘露醇mannitol | 31.9±2.4 cB | 44.7±1.5 cA | 44.5±2.3 bA | 32.5±3.2 dB |
海藻糖trehalose | 11.2±1.6 dB | 22.3±2.6 dA | 18.7±3.9 cA | 19.7±1.3 eA |
表2
香樟体胚诱导率的影响因素回归分析"
因素类型 factor type | factor 系数 coefficient | 标准差 standard deviation | 标准化系数 standard coefficient | t | P |
---|---|---|---|---|---|
常量constant | 67.786 | 7.339 | 9.236 | <0.001*** | |
产地different original location | -0.425 | 2.055 | -0.020 | -0.207 | 0.837 |
碳源种类types of carbon sources | -8.251 | 1.346 | -0.594 | -6.132 | <0.001*** |
整体模型integral model | R2=0.353 F (2,69) =18.82 | R2=0.334 P<0.001*** |
表3
两种混合碳源对长沙产地的香樟体细胞胚诱导的影响"
混合碳源质量浓度/(g·L-1) concentration of two mixed carbon sources | 体胚诱导率/% induction rate of somatic embryo | ||
---|---|---|---|
蔗糖 source | 山梨醇 sorbitol | 甘露醇 mannitol | |
45(CK) | 84.1±1.3 c | ||
15 | 15 | 87.7±2.3 b | |
15 | 30 | 92.9±1.7 a | |
15 | 45 | 55.3±1.4 d | |
15 | 60 | 31.9±2.5 e | |
45(CK) | 84.1±1.3 b | ||
15 | 15 | 85.4±1.7 b | |
15 | 30 | 90.8±1.1 a | |
15 | 45 | 55.1±3.2 c | |
15 | 60 | 30.7±2.5 d |
表6
两种混合碳源对长沙产地的香樟植株再生的影响"
两种混合碳源质量浓度/(g·L-1) concentration of two mixed carbon sources | 体胚芽 萌发率/% bud germination rate | 体胚 生根率/% rooting rate | ||
---|---|---|---|---|
蔗糖 sucrose | 山梨醇 sorbitol | 甘露醇 mannitol | ||
45(CK) | 53.3±1.2 a | 85.4±4.2 a | ||
15 | 15 | 54.7±1.7 a | 86.9±3.7 a | |
15 | 30 | 47.3±1.7 b | 84.3±3.5 a | |
15 | 45 | 47.2±3.6 b | 84.2±3.2 a | |
45(CK) | 53.3±1.2 a | 85.4±4.2 a | ||
15 | 15 | 53.6±2.7 a | 86.0±2.3 a | |
15 | 30 | 46.8±1.6 b | 83.9±1.6 a | |
15 | 45 | 41.9±2.6 c | 64.4±2.3 b |
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