Induction of pollen embryo and chromosome doubling in tobacco (Nicotiana tabacum L.)
Another culture is an effective breeding method for improving desirable traits, since it allows fast achievement of genetically homozygous lines. The present studies were carried out to evaluate the response of flue-cured tobacco anthers for high frequency pollen embryo induction in different culture media, and the effects of colchicine treatment on tobacco chromosome doubling. The results showed that different media caused significant changes in pollen embryo induction, in which the most obvious change was found in B medium with induction rates of 23.28%, 26.64%, and 28.07% for TMK-12 × Jiyan 5, K326 × Shangkang 70, and NC71 × Jingyehuang, respectively. In addition, shoot tips of the haploid aseptic seedlings cultivated from anthers were treated with 4 g/L colchicine for 48 h, and the ploidy levels were then checked by using flow cytometry and the number of chloroplasts in stomatal guard cells. The results suggested that colchicine treatment not only doubled the chromosomes of sterile haploid seedlings, but also caused some sterile haploid seedlings to die. The survival rate of TMK-12 × Jiyan 5, K326 × Shuangkang 70, and NC71 × Jingyehuang seedlings were 33.29%, 35.11%, and 17.61%, respectively. Similarly, TMK-12 × Jiyan 5 and K326 × Shuangkang 70 also showed a relatively high chromosome doubling rate compared with NC71 × Jingyehuang; their chromosome doubling rates were 21.53%, 18.54%, and 7.98%, respectively. In general, our work provides a theoretical basis for accelerating the breeding and improvement of new flue-cured tobacco varieties.
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- Abdoli M, Moieni A, Badi HN (2013). Morphological, physiological,
cytological and phytochemical studies in diploid and
colchicine-induced tetraploid plants of Echinacea purpurea
(L.). Acta Physiologiae Plantarum 35(7): 2075-2083. doi:
10.1007/s11738-013-1242-9
- Bagheri M, Mansouri H (2015). Effect of induced polyploidy on
some biochemical parameters in Cannabis sativa L. Applied
Biochemistry and Biotechnology 175(5): 2366-2375. doi:
10.1007/s12010-014-1435-8
- Beck S, Visser G, Dunlop R, Hare P (2005). A comparison of direct
(flow cytometry) and indirect (stomatal guard cell lengths and
chloroplast numbers) techniques as a measure of ploidy in black
wattle, Acacia mearnsii (de Wild). South African Journal of
Botany 71(3-4): 354-358. doi: 10.1016/S0254-6299(15)30109-5
- Burk LG, Gwynn GR, Chaplin JF (1972). Diploidized haploids from
aseptically cultured anthers of Nicotiana tabacum: A colchicine
method applicable to plant breeding. Journal of Heredity 63(6):
355-360. doi: 10.1093/oxfordjournals.jhered.a108316
- Chen XJ, Peng SY, Luo JR, Yang YM, Xiao BG (2011). Culture of
regenerated seedlings from anthers and construction of DH
populations of six cross combinations of Nicotiana tabacum,
Journal of Plant Resources and Environment 20(1): 65-68 (in
Chinese with an abstract in English). doi: 10.3969/j.issn.1674-
7895.2011.01.010
- Choudhuri HC, Banerjee S (1961). Colchicine treatment of different
varieties of potatoes in West Bengal. European Potato Journal
4(2): 144-151. doi: 10.1007/BF02365026
- Clarkson JJ, Chase MW, Knapp S (2004). Nomenclatural changes and
a new sectional classification in Nicotiana (Solanaceae). Taxon
53(1): 73-82. doi: 10.2307/4135490
- Dolezel J, Greilhuber J, Suda J (2007). Estimation of nuclear DNA
content in plants using flow cytometry. Nature Protocols 2(9):
2233-2244. doi: 10.1038/nprot.2007.310
- Dunwell JM (2010). Haploids in flowering plants: origins and
exploitation. Plant Biotechnology Journal 8(4): 377-424. doi:
10.1111/j.1467-7652.2009.00498.x
- Forster BP, Heberle-Bors E, Kasha KJ, Touraev A (2007). The
resurgence of haploids in higher plants. Trends in Plant Science
12(8): 368-375. doi: 10.1016/j.tplants.2007.06.007
- Forster BP, Thomas WTB (2005). Doubled haploids in genetics
and plant breeding. Plant Breeding Reviews 25: 57-88. doi:
10.1002/9780470650301.ch3
- Germanà MA (2006). Doubled haploid production in fruit crops.
Plant Cell Tissue and Organ Culture 86(2): 131-146. doi:
10.1007/s11240-006-9088-0
- Germanà MA (2011). Anther culture for haploid and doubled
haploid production. Plant Cell Tissue and Organ Culture
104(3): 283-300. doi: 10.1007/s11240-010-9852-z
- Giannelos PN, Zannikos F, Stournas S, Lois E, Anastopoulos G
(2002). Tobacco seed oil as an alternative diesel fuel: physical
and chemical properties. Industrial Crops and Products 16(1):
1-9. doi: 10.1016/S0926-6690(02)00002-X
- Hamid HM, Ghasem K, Reza D, Reza NM, Ahmad S (2013).
Identification of QTLs associated with low chloride
accumulation in oriental tobacco. Genetika 45(3): 855-864.
doi: 10.2298/GENSR1303855M
- Heberle-Bors E (1980). Interaction of activated charcoal and iron
chelates in anther cultures of Nicotiana and Atropa belladonna.
Zeitschrift für Pflanzenphysiologie 99(4): 339-347. doi:
10.1016/S0044-328X(80)80148-6
- Herath H, Bandara DC, Samarajeewa PK, Wijesundara DSA (2009).
Effect of low temperature pre-treatment on anther culture in
selected indica, japonica rice varieties and their inter subspecific hybrids. Ceylon Journal of Science, Biological Sciences
38(1): 11-16. doi: 10.4038/cjsbs.v38i1.1322
- Jaskani MJ, Kwon SW, Dae A, Kin H (2005). Flow cytometry of DNA
contents of colchicine treated watermelon as a ploidy screening
method at M1 stage. Pakistan Journal of Botany 37(3): 685-
696. doi: 10.1111/j.1469-8137.2005.01437.x
- Kumar HGA, Murthy HN (2004). Effect of sugars and amino
acids on androgenesis of Cucumis sativus. Plant Cell Tissue
and Organ Culture 78(3): 201-208. doi: 10.1023/B:TI
CU.0000025637.56693.68
- Labbani ZJ. De Buyser, Picard E (2010). Effect of mannitol
pretreatment to improve green plant regeneration on isolated
microspore culture in Triticum turgidum ssp. durum cv. ‘Jennah
Khetifa’. Plant Breeding 126(6): 565-568. doi: 10.1111/j.1439-
0523.2007.01399.x
- Li H, Yan B, Zhang T, Jiang YL, Zhang H et al. (2009). Preliminary
studies on polyploidy mutation of cut flower Gerbera
jamesonii Bolus. Acta Horticulturae Sinica 36(4): 605-610 (in
Chinese with an abstract in English). doi: 10.3321/j.issn:0513-
353X.2009.04.023
- Li Y, Li H, Chen Z, Ji LX, Ye MX et al. (2013). Haploid plants from
anther cultures of poplar (Populus × beijingensis). Plant Cell
Tissue and Organ Culture 114(1): 39-48. doi: 10.1007/s11240-
013-0303-5
- Liu R, Jiang G, Yang S, Huang N (2009). Study on the double effect of
colchicine concentration and seedling dipping time on tobacco
haploid seedling. Seed 28(1): 12-15 (in Chinese with an
abstract in English). doi: 10.3969/j.issn.1001-4705.2009.01.004
- Lu R, Chen Z, Gao R, He T, Wang Y et al. (2016). Genotypesindependent optimization of nitrogen supply for isolated
microspore cultures in barley. Biomed Research International
2016: 1-8. doi: 10.1155/2016/1801646
- Mitykó J, Andrásfalvy A, Csilléry G, Fári M (2010). Anther-culture
response in different genotypes and F1
hybrids of pepper
(Capsicum annuum L.). Plant Breeding 114(1): 78-80. doi:
10.1111/j.1439-0523.1995.tb00764.x
- Nakamura A, Takashima S, Hasegawa H, Matsumoto T (2008).
Simple and efficient chromosome doubling method by
colchicine-treatment for haploid plantlets of tobacco. Breeding
Science 43(4): 603-612. doi: 10.1270/jsbbs1951.43.603
- Nitsch C, Norreel B (1973). Factors favoring the formation of
androgenetic embryos in anther culture. Basic Life Sciences
2(11): 129-144. doi: 10.1007/978-1-4684-2880-3-10
- Nitsch JP, Nitsch C (1969). Haploid plants from pollen grains.
Science 163: 85-87. doi: 10.1126/science.163.3862.85
- Ojiewo CO, Agong SG, Murakami K, Masuda M (2006).
Chromosome duplication and ploidy level determination
in African nightshade Solanum villosum Miller. Journal of
Pomology and Horticultural Science 81(2): 183-188. doi:
10.1080/14620316.2006.11512048
- Peng J, Song K, Zhu H, Kong W, Liu F (2017). Fast detection of
tobacco mosaic virus infected tobacco using laser-induced
breakdown spectroscopy. Scientific Reports 7: 44551. doi:
10.1038/srep44551
- Ramulu KS, Verhoeven HA, Dijkhuis P (1991). Mitotic blocking,
micronucleation, and chromosome doubling by oryzalin,
amiprophos-methyl, and colchicine in potato. Protoplasma
160(2-3): 65-71. doi: 10.1007/BF01539957
- Redha A, Attia T, Buter B, Stamp P, Schmid JE (1998). Single and
combined effects of colchicine, L-proline and post-inoculation
low temperature on anther culture of wheat, Triticum aestivum
L. Plant Breeding 117(4): 335-340. doi: 10.1111/j.1439-
0523.1998.tb01950.x
- Salas P, Prohens J, José M (2011). Evaluation of androgenic
competence through anther culture in common eggplant
and related species. Euphytica 182(2): 261-274. doi: 10.1007/
s10681-011-0490-2
- Sato S, Katoh N, Iwai S, Hagimori M (2002). Effect of low temperature
pretreatment of buds or inflorescence on isolated microspore
culture in Brassica rapa (syn. B. campestris). Breeding Science
52(1): 23-26. doi: 10.1270/jsbbs.52.23
- Seguı´-Simarro JM, Nuez F (2008). How microspores transform
into haploid embryos: changes associated with embryogenesis
induction and microspore-derived embryogenesis. Physiologia
Plantarum 134(1): 1-12. doi: 10.1111/j.1399-3054.2008.01113.x
- Song H, Lou QF, Luo XD, Wolukau JN, Diao WP et al. (2007).
Regeneration of doubled haploid plants by androgenesis of
cucumber (Cucumis sativus L.). Plant Cell Tissue and Organ
Culture 90(3): 245-254. doi: 10.1007/s11240-007-9263-y
- Sopory SK, Maheshwari SC (1976). Development of pollen
embryoids in anther cultures of Datura innoxia ii. effects of
growth hormones. Journal of Experimental Botany 27(1): 58-
68. doi: 10.1093/jxb/27.1.58
- Soriano M, Cistue L, Valles MA (2007). Effects of colchicine on
anther and microspore culture of bread wheat (Triticum
aestivum L.). Plant Cell Tissue and Organ Culture 91(3): 225-
234. doi: 10.1007/s11240-007-9288-2
- Soriano M, Li H, Boutilier K (2013). Microspore embryogenesis:
establishment of embryo identity and pattern in culture. Plant
Reproduction 26(3): 181-196. doi: 10.1007/s00497-013-0226-7
- Sunderland N, Roberts M (1979). Cold-pretreatment of excised
flower buds in float culture of tobacco anthers. Annals of
Botany 43(4): 405-414. doi: 10.2307/2442222
- Takashima S, Hasegawa H, Nakamura A (1995). A simple method
for chromosome doubling in tobacco anther culturedirect application of colchicine to anthers before culture.
Japanese Journal of Breeding 45(1): 107-110. doi: 10.1270/
jsbbs1951.45.107
- Tavan M, Mirjalili MH, Karimzadeh G (2015). In vitro polyploidy
induction: changes in morphological, anatomical and
phytochemical characteristics of Thymus persicus (Lamiaceae).
Plant Cell Tissue and Organ Culture 122(3): 573-583. doi:
10.1007/s11240-015-0789-0
- Tazawa M, Reinert J (1969). Extracellular and intracellular
chemical environments in relation to embryogenesis in vitro.
Protoplasma 68(1-2): 157-173. doi: 10.1007/BF01247902
- Vardar F, Aydin Y, Ayturk O, Unal M (2017). The Effects of colchicine
on growth and development of sunflower (Helianthus annuus
L) seedlings. Fresenius Environmental Bulletin 17: 4113-4118.
- Wetherell DF, Dougall DK (2006). Sources of nitrogen supporting
growth and embryogenesis in cultured wild carrot tissue.
Physiologia Plantarum 37(2): 97-103. doi: 10.1111/j.1399-
3054.1976.tb03939.x
- Zhang J, Li G, Qiao Y, Wang M, Li J et al. (2009). Effects of colchicine
on regeneration and polyploid induction from leaf in vitro of
strawberry (Fragaria × ananassa). Journal of Plant Resources
and Environment 18(3): 69-73 (in Chinese with an abstract in
English). doi: 10.3969/j.issn.1674-7895.2009.03.011
- Zhao J, Zhou XX, Zhang ZQ, Yang BZ, Zhou SD (2010). Effects of
culture media on anther culture of chili pepper (Capsicum
annuum L.). Journal of Hunan Agricultural University
(Natural Sciences) 36(2): 181-187 (in Chinese with an abstract
in English). doi:10.3724/SP.J.1238.2010.00181
- Zhao L, Hu R, Cai C, Zhou Z, Liu Z et al. (2015). Influence of
colchicines on chromosome doubling of tobacco haploid.
Crop Research 29(3): 235-239 (in Chinese with an abstract in
English). doi: 10.3969/j.issn.1001-5280.2015.03.04
- Zhou K, Fleet P, Nevo E, Zhang X, Sun G (2017). Transcriptome
analysis reveals plant response to colchicine treatment during
on chromosome doubling. Scientific Reports 7(1): 8503. doi:
10.1038/s41598-017-08391-2
- Zhou ZG, Gong YQ, Wang XW, Liu LW, Zhang YG (2007). Induction
of isolated microspore and optimization of their culture system
of different radish varieties. Acta Botanica Boreali–Occidentalia
Sinica 27(1): 33-38. doi: 10.1016/S1872-2075(07)60067-3