Abstract
Thuarea involuta, a perennial herb that often grows on sandy beaches, has documented salt and drought tolerance, making it a useful plant species for the stabilization of island reefs. However, there are no protocols for its large-scale propagation, so effective in vitro protocols can assist to mass propagate healthy plants for ecorestoration. In this study, T. involuta stem segments were used as explants to establish a highly efficient in vitro shoot proliferation and regeneration system for the first time. The shoot proliferation coefficient (SPC) was highest when thidiazuron was used alone (SPC = 8.78) relative to the use of kinetin (KIN) or 6-benzyladenine (SPC = 6.53 and 8.30, respectively). Multiple shoots were proliferated on Murashige and Skoog (MS) medium supplemented with 1.0 mg L−1 BA and 0.1 mg L−1 α-naphthaleneacetic acid, leading to a SPC of 9.64 within 30 days. After individual shoots were transferred to MS medium with or without indole-3-butyric acid (IBA) or α-naphthaleneacetic acid (NAA), all formed roots within 15 days. In vitro plantlets transplanted to a peat and vermiculite (1:1) substrate displayed 96.67% survival after 60 days. When MS medium was supplemented with 1.0 mg L−1 2,4-dichlorophenoxyacetic acid and 1.15 g L−1 proline, callus was effectively induced from the base of shoots; 90% of callus differentiated into adventitious shoot buds on MS medium containing 0.1 mg L−1 KIN. This study provides a useful set of in vitro protocols for the practical mass propagation of plants that can be used in sand dune ecorestoration, or to avoid the erosion of beach sand by sea water.
Key message
An efficient in vitro shoot proliferation, callus induction, differentiation, plant regeneration and transplantation system in a salt-tolerant plant Thuarea involuta was established for the first time.
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Data availability
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Abbreviations
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- BA:
-
6-Benzyladenine
- IBA:
-
Indole-3-butyric acid
- KIN:
-
Kinetin
- MS:
-
Murashige and Skoog
- NAA:
-
α-Naphthaleneacetic acid
- PGR:
-
Plant growth regulator
- SPC:
-
Shoot proliferation coefficient
- TDZ:
-
Thidiazuron
References
Abdelgawad H, Zinta G, Hegab MM, Renu P, Han A, Walid A (2016) High salinity induces different oxidative stress and antioxidant responses in maize seedlings organs. Front Plant Sci 7:276. https://doi.org/10.3389/fpls.2016.00276
Chai M, Jia Y, Chen S, Gao ZS, Wang HF, Liu LL, Wang PJ, Hou DQ (2011) Callus induction, plant regeneration, and long-term maintenance of embryogenic cultures in Zoysia matrella [L.] Merr. Plant Cell Tiss Org Cult 104:187–192. https://doi.org/10.1007/s11240-010-9817-2
Chen YL, Li X, Zhao WZ, Li XJ, Liao HM, Liu DM (2022) Physiological response of Thuarea involuta under salt stress. Guihaia 41:225–232. https://doi.org/10.11931/guihaia.gxzw202007040 (in Chinese with English abstract)
Fatima S, Mujib A, Samaj J (2011) Anti-oxidant enzyme responses during in vitro embryogenesis in Catharanthus roseus. J Horticult Sci Biotech 86:569–574. https://doi.org/10.1080/14620316.2011.11512805
Giri CC, Praveena M (2015) In vitro regeneration, somatic hybridization and genetic transformation studies: an appraisal on biotechnological interventions in grasses. Plant Cell Tiss Org Cult 120:843–860. https://doi.org/10.1007/s11240-014-0653-7
Griffin JD, Dibble MS (1995) High-frequency plant regeneration from seed-derived callus cultures of Kentucky bluegrass (Poa pratensis l.). Plant Cell Rep 14:721–724. https://doi.org/10.1007/BF00232655
Guo B, Abbasi BH, Zeb A, Xu LL, Wei YH (2011) Thidiazuron: a multi-dimensional plant growth regulator. Afr J Biotech 10:8984–9000. https://doi.org/10.1186/1471-2164-12-418
Hayasaka D, Goka K, Thawatchai W (2012) Ecological impacts of the 2004 Indian Ocean tsunami on coastal sand-dune species on Phuket Island, Thailand. Biodivers Conserv 21:1971–1985. https://doi.org/10.1007/s10531-012-0288-0
Levin MJ (2019) Prehistoric agricultural transformations in tropical remote Oceania: towards comprehensive phytolith analysis. Quat Intern 529:70–95. https://doi.org/10.1016/j.quaint.2019.03.002
Li L, Qu R (2004) Development of highly regenerable callus lines and biolistic transformation of turf-type common bermudagrass (Cynodon dactylon L. Pers.). Plant Cell Rep 22:403–407. https://doi.org/10.13989/j.cnki.0517-6611.2009.29.113
Liu L, Fang XL, Zhang JW, Bao YM, Yan M (2009) Long-term cultured callus and the effect factor of high-frequency plantlet regeneration and somatic embryogenesis maintenance in Zoysia japonica. In Vitro Cell Dev Biol—Plant 45:673–680. https://doi.org/10.1007/s11627-009-9226-6
Lu SY, Wang ZC, Peng XX, Guo Z, Zhang G, Han L (2006) An efficient callus suspension culture system for triploid Bermuda grass (Cynodon transvaalensis × C. dactylon) and somaclonal variations. Plant Cell Tiss Org Cult 87:77–84. https://doi.org/10.1007/s11240-006-9138-7
Ma GH, Jian SG, Ren H (eds) (2021) Thuarea involuta (G. Forst.) R. Br. ex Sm. In: Plant Proliferation and Cultivation Techniques for Tropical Island Reefs. China Forestry Press, Bei**g, pp 6–8. ISBN: 978-7-5219-0955-5 (in Chinese)
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Nanjo T, Kobayashi M, Yoshiba Y, Sanada Y, Wada K, Tsukaya H, Kakubari Y, Yamaguchi-Shinozaki K, Shinozaki K (1999) Biological functions of proline in morphogenesis and osmotolerance revealed in antisense transgenic Arabidopsis thaliana. Plant J 18:185–193. https://doi.org/10.1046/j.1365-313X.1999.00438.x
Neibaur I, Altpeter GF (2008) The effect of auxin type and cytokinin concentration on callus induction and plant regeneration frequency from immature inflorescence segments of seashore paspalum (Paspalum vaginatum Swartz). In Vitro Cell Dev Biol - Plant 44:480–486. https://doi.org/10.1007/s11627-008-9143-0
Plentovich S, Mizerek T, Reeves MK, Amidon F, Nanbara M (2020) Coastal strand and mangrove swamps of the Mariana Islands. Encycl World’s Biomes 1:185–197. https://doi.org/10.1016/B978-0-12-409548-9.12422-4
Reinheimer R, Vegetti AC (2008) Inflorescence diversity and evolution in the PCK clade (Poaceae: Panicoideae: Paniceae). Plant Syst Evol 275:133–167. https://doi.org/10.3732/ajb.0800245
Ren H, Jian SG, Zhang QM. Wang FG, Shen T, Wang J (2017) Plants and vegetation on South China Sea Islands. Ecol Environ Sci 26:1639–1648. http://www.jeesci.com (in Chinese with English abstract)
Sarabi B, Bolandnazar S, Ghaderi N, Ghashghaie J (2017) Genotypic differences in physiological and biochemical responses to salinity stress in melon (Cucumis melo L.) plants: prospects for selection of salt tolerant landraces. Plant Physiol Biochem 119:294–311. https://doi.org/10.1016/j.plaphy.2017.09.006
Wang Y, Ruemmele BA, Chandlee JM, Chandlee JM, Sullivan WM, Knapp JE, Kausch AP (2002) Embryogenic callus induction and plant regeneration media for bentgrasses and annual bluegrass. In Vitro Cell Dev Biol—Plant 38:460–467. https://doi.org/10.1079/IVP2002337
Woo HS, Lee SH, Lee DG, Kim JS, Lee BH (2004) Efficient plant regeneration from mature seed-derived callus of Italian ryegrass (Lolium multiflorum Lam.). Kor J Plant Biotech 31:43–48. https://doi.org/10.5010/JPB.2004.31.1.043
Yuan X, Wang Z, Liu J, She J (2009) Development of a plant regeneration system from seed-derived calluses of centipedegrass [Eremochloa ophiuroides (munro.) hack]. Sci Hortic 120:96–100. https://doi.org/10.1016/j.scienta.2008.09.005
Zhang WJ, Dong JL, Liang BG, Wang JT (2006) Highly efficient embryogenesis and plant regeneration of tall fescue (Festuca arundinacea Schreb.) from mature seed-derived calli. In Vitro Cell Dev Biol—Plant 42:114–118. https://doi.org/10.1079/IVP2005733
Zhang S, Hanna W, Ozias-Akins P (2007) Comparison of callus induction and plant regeneration from different explants in triploid and tetraploid turf-type Bermuda grasses. Plant Cell Tiss Org Cult 90:71–78. https://doi.org/10.1007/s11240-007-9255-y
Zhao K, Hai F, Ungar IA (2002) Survey of halophyte species in China. Plant Sci 163:491–498. https://doi.org/10.1016/S0168-9452(02)00160-7
Funding
This work was financially supported by National Key Research & Development Program of China (2021YFC3100400, 2022YFC3103700) and Guangdong Key Areas Biosafety Project (2022B1111040003).
Author information
Contributions
ZPW and YPX designed the experiment and provided guidance for the study. YJZ and JYL prepared samples for all analyses. ZPW, KLW, SJZ and YJZ conducted the experiments and statistical analyses. JATS provided advice, interpretation of the experiment and analyses, and co-wrote the manuscript with GHM. All authors read and approved the manuscript for publication.