Plant Cell, Tissue and Organ Culture (PCTOC) (2018) 132:425–432
In vitro induction and identiﬁcation of autotetraploid of Bletilla striata
(Thunb.) Reichb.f. by colchicine treatment
· Liu Wei‑xu
· Liang Hui‑hui
· Xiang zeng‑xu
Received: 25 July 2017 / Accepted: 15 October 2017 / Published online: 21 November 2017
© Springer Science+Business Media B.V., part of Springer Nature 2017
Polyploidy breeding has proved to be a valuable approach for acquiring the high yield superior varieties in medicinal plants.
An eﬀective protocol for obtaining Bletilla striata autotetraploid is in vitro induction of protocorms with colchicine. The
protocorms of B.striata were soaked in diﬀerent concentrations of colchicine solution [0.05, 0.1 and 0.2% (w/v)] for 12,
24, 36, 48 and 60 h, and the ploidy of the seedlings was identiﬁed by chromosome counting and ﬂow cytometry analysis.
The results showed that the optimal condition for induction of autotetraploid of B. striata protocorms was treated with 0.2%
colchicine for 36 h with the induction rate reached as high as 26.7%. In addition, the morphological and anatomical charac-
teristics were observed and compared between the diploid and tetraploid plants. And we found that the features of tetraploid
plants were signiﬁcantly diﬀerent from diploid plants, such as tetraploid plants possessed thicker and deeper green leaves,
larger stomata and more chloroplast number, which could be used as simple and eﬃcient parameters for screening tetraploid.
This study laid a foundation for breeding superior varieties of B. striata.
Keywords Bletilla striata · In vitro induction · Colchicine · Autotetraploid
Bletilla striata (Thunb.) Reichb.f. is an endangered orchid
with high ornamental and medicinal value. The dry tuber
of B. striata is a traditional Chinese medicine which has
been widely used for thousands of years. B. striata has mul-
tiple eﬀects including hemostasis with astringents, detu-
mescence and promoting granulation (Commission 2015).
Recently, several researches indicate that the polysaccharide
is the main eﬀective component of B. striata which plays
an important role in the treatment of tumors, pneumococ-
cal pneumonia, and wound healing (Bonten et al. 2015; Lv
et al. 2012; Pereira et al. 2016; Wang et al. 2011). Polysac-
charide in B. striata possesses obvious curative eﬀects on
postponing cutaneous caducity, curing or preventing cutane-
ous chilblain and chap, accelerating blood cycle and improv-
ing cutaneous nutrition status without reported side eﬀect
(Sun et al. 2009). Polysaccharide has been used as a natural
thickener, suspending agent, moisturizer and emulsiﬁer in
cosmetic industries (Sun et al. 2009). Therefore, the extrac-
tion of higher content and better quality of polysaccharide
from B. striata could increase the economic value.
In recent years, the demand for natural herbal was
increased with the rapid development of traditional Chi-
nese medicine and improvement of people’s living stand-
ards. However, the limited wild resources, unrestricted and
unplanned excavation of medicinal plants and extensive dis-
turbance of their natural habitat make it facing a shortage
crisis (Tu 2006). Besides, the yield and quality of medicinal
material have been reduced because of the degradation of
cultivated varieties in the process of long-term cultivation
(Zhibin 2008). Therefore, it is necessary to breed supe-
rior varieties of medicinal plants with good quality, high
yield, more secondary metabolites, broad and high level of
Communicated by Sergio J. Ochatt.
* Xiang zeng-xu
Institute of Chinese Medicinal Materials, Nanjing
Agricultural University, No. 1 Weigang Road, Nanjing,
Jiangsu 210095, China