In vitro Germination,Callus Induction and Primary Haustorium Organogenesis in the Parasitic Plant Cynomorium songaricum

doi:10.7525/j.issn.1673-5102.2020.06.007

Abstract

Abstract:

Cynomorium songaricum Rupr. is an obligate parasitic plant widely used as traditional Chinese medicine and Mongolian medicine. Here， we firstly describe protocols for in vitro germination， callus induction and haustorium organogenesis in C.songaricum Rupr. In this study， adequate concentrations of gibberellic acid（GA₃），_{in combination with other plant growth regulators in the medium， promoted embryo development and germination of C.songaricum seeds. The highest callus induction rate from seeds（13.7%） after a 40 d incubation was obtained with B₅ medium adding 2，4-dichlorophenoxyacetic acid（2，4-D； 1.0 mg·L^-1）， kinetin（KT；0.5 mg·L^-1） and GA₃（1.0 mg·L^-1. This resulted in callus_{formation in 13.7%±3.1% of seeds. Addition of 2，4-D（0.5 mg·L^-1） and KT（0.25 mg·L^-1） yielded highest haustorium organogenesis from calluses. Some primary haustorium branched to form adventitious roots of 3-4 cm in length. Subsequently， the tip of each adventitious root formed a nascent primary haustorium， which was then branched out into adventitious roots. The role of auxins（2，4-D in this study） in the formation of primary haustorium and adventitious roots from seed callus in C.songaricum was also discussed.}}

Key words: medicinal plant, HIFs, histology, host-parasite relationship, cultivation

CLC Number:

Q949.762.9

Xin YUE, Gui-Lin CHEN. In vitro Germination,Callus Induction and Primary Haustorium Organogenesis in the Parasitic Plant Cynomorium songaricum[J]. Bulletin of Botanical Research, 2020, 40(6): 846-854.

Figures/Tables 5

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References 36

1	Jin S W，Eerdunbayaer，Doi A，et al.Polyphenolic constituents of Cynomoriumsongaricum Rupr.and antibacterial effect of polymeric proanthocyanidin on methicillin-resistant Staphylococcusaureus［J］.Journal of Agricultural and Food Chemistry，2012，60（29）：7297-7305.
2	Ma C M，Nakamura N，Miyashiro H，et al.Inhibitory effects of constituents from Cynomoriumsongaricum and related triterpene derivatives on HIV-1 protease［J］.Chemical and Pharmaceutical Bulletin，1999，47（2）：141-145.
3	Ma C M，Wei Y，Wang Z G，et al.Triterpenes from Cynomoriumsongaricium-analysis of HCV protease inhibitory activity，quantification，and content change under the influence of heating［J］.Journal of Natural Medicines，2009，63（1）：9-14.
4	Nakamura N.Inhibitory effects of some traditional medicines on proliferation of HIV-1 and its protease［J］.Yakugaku Zasshi，2004，124（8）：519-529.
5	Press M C，Scholes J D，Watling J R.Parasitic plants：physiological and ecological interactions with their hosts［M］.//Press M C，Scholes J D，Barker M G.Physiological Plant Ecology.Cambridge：Cambridge University Press，1999：175-97.
6	Yoshida S，Shirasu K.Plants that attack plants：molecular elucidation of plant parasitism［J］.Current Opinion in Plant Biology，2012，15（6）：708-713.
7	Keyes W J，Taylor J V，Apkarian R P，et al.Dancing together.Social controls in parasitic plant development［J］.Plant Physiology，2001，127（4）：1508-1512.
8	Shen H，Ye W，Hong L，et al.Progress in parasitic plant biology：host selection and nutrient transfer［J］.Plant Biology，2006，8（2）：175-185.
9	Zehhar N，Ingouff M，Bouya D，et al.Possible involvement of gibberellins and ethylene in Orobancheramosa germination［J］.Weed Research，2010，42（6）：464-469.
10	Mohamed A H，Ejeta G，Housley T L.Strigaasiatica seed conditioning and 1-aminocyclopropane-1-carboxylate oxidase activity［J］.Weed Research，2010，41（2）：165-176.
11	Stewart G R，Press M C.The physiology and biochemistry of parasitic angiosperms［J］.Annual Review of Plant Physiology and Plant Molecular Biology，1990，41：127-151.
12	Yoshida S，Cui S K，Ichihashi Y，et al.The Haustorium，a specialized invasive organ in parasitic plants［J］.Annual Review of Plant Biology，2016，67：643-667.
13	Spallek T，Melnyk C W，Wakatake T，et al.Interspecies hormonal control of host root morphology by parasitic plants［J］.Proceedings of the National Academy of Sciences of the United States of America，2017，114（20）：5283-5288.
14	Keyes W J，O'malley R C，Kim D，et al.Signaling organogenesis in parasitic angiosperms：Xenognosin generation，perception，and response［J］.Journal of Plant Growth Regulation，2000，19（2）：217-231.
15	Ihl B，Jacob F，Sembdner G.Studies on Cuscutareflexa ROXB.V.The level of endogenous hormones in the parasite，Cuscuta reflexa，and its host，Viciafaba L.，and a suggested role in the transfer of nutrients from host to parasite［J］.Plant Growth Regulation，1984，2（2）：77-90.
16	Klaren C H.Physiological aspects of the hemiparasite Rhinanthus serotinus.Groningen：University of Groningen，1975.
17	Paliyath G，Maheshwari R，Mahadevan S.Initiation of Haustoria in Cuscuta by Cytokinin application［J］.Current Science，1978，47（12）：427-429.
18	Huang J Z，Li Y H.A study on the developmental anatomy of haustoria in Cusutajaponca choisy induced by exogenous cytokinin application［J］.Journal of Nanjing Agricultural University，1991，14（2）：23-28.
19	Dong S L，Chen N L，Zhang Y，et al.Preliminary study on dormancy and germination of Cynomoriumsongaricum seed［J］.Arid Zone Research，2011，28（4）：654-659.
20	Mwase W，Mvula T.Effect of seed size and pre-treatment methods of Bauhiniathonningii Schum.on germination and seedling growth［J］.African Journal of Biotechnology，2011，10（26）：5143-5148.
21	Bewley J D.Seed germination and dormancy［J］.Plant Cell，1997，9（7）：1055-1066.
22	Ren M Y，Yang G，Du L S，et al.Research advances in medicinal plants of Cynomorium songaricum［J］.Journal of Biology，2018，35（5）：95-98.
23	Joel D M，Back A，Kleifeld Y，et al.Seed conditioning and its role in Orobanche seed germination：inhibition by paclobutrazol［M］.//Wegmann K，Musselman L J.Progress in Orobanche Research.Proceedings of the International Workshop on Orobanche Research.Tubingen：FRG，1991：147-156.
24	Joel D M，Stevens J C，Matthews D E.Germination of weedy root parasites［M］.//Kigel J，Galili C.Seed Development and Germination.New York：Marcel Dekker，1995：567-598.
25	Joel D M.The long-term approach to parasitic weeds control：manipulation of specific developmental mechanisms of the parasite［J］.Crop Protection，2000，19（8-10）：753-758.
26	Deeks S J，Shamoun S F，Punja Z K.In vitro germination and development of western hemlock dwarf mistletoe［J］.Plant Cell，Tissue and Organ Culture，2001，66（2）：97-105.
27	Ang S L P，Yong J W H.A protocol for invitro germination and sustainable growth of two tropical mistletoes［J］.Plant Cell，Tissue and Organ Culture，2005，80（2）：221-228.
28	Das P，Kar M，Sahoo S.In vitro hormone-regulated growth and ﬂoral induction of Cuscutareﬂexa：a parasitic angiosperm［J］.Acta Physiologiae Plantarum，2011，33（3）：1031-1035.
29	Cui S K，Wada S，Tobimatsu Y，et al.Host lignin composition affects haustorium induction in the parasitic plants Phtheirospermumjaponicum and Strigahermonthica［J］.New Phytologist，2018，218（2）：710-723.
30	Tomilov A A，Tomilova N B，Abdallah I，et al.Localized hormone fluxes and early haustorium development in the hemiparasitic plant Triphysariaversicolor［J］.Plant Physiology，2005，138（3）：1469-1480.
31	Kuijt J.The biology of parasitic flowering plants［M］.Berkeley：University of California Press，1969.
32	Ishida J K，Yoshida S，Ito M，et al.Agrobacterium rhizogenes-mediated transformation of the parasitic plant Phtheirospermumjaponicum［J］.PLoS One2011，6（10）：e25802.
33	Bandaranayake P C G，Filappova T，Tomilov A，et al.A single-electron reducing quinone oxidoreductase is necessary to induce haustorium development in the root parasitic plant Triphysaria［J］.Plant Cell，2010，22（4）：1404-1419.
34	Kokla A，Melnyk C W.Developing a thief：Haustoria formation in parasitic plants［J］.Developmental Biology，2018，442（1）：53-59.
35	Goyet V，Billard E，Pouvreau J B，et al.Haustorium initiation in the obligate parasitic plant Phelipancheramosa involves a host-exudated cytokinin signal［J］.Journal of Experimental Botany，2017，68（20）：5539-5552.
36	Ishida J K，Wakatake T，Yoshida S，et al.Local auxin biosynthesis mediated by a YUCCA flavin monooxygenase regulates Haustorium development in the parasitic plant Phtheirospermumjaponicum［J］.Plant Cell，2016，28（8）：1795-1814.

Treatment	Plant growth regulator(mg·mL^-1)			Rate of callus induction(%)
Treatment	2,4-D	KT	GA₃	Rate of callus induction(%)
1	0.5	0.1	0.5	1.7±1.5^f
2	1.0	0.5	1.0	13.7±3.1^a
3	2.0	1.0	2.0	9.0±1.0^b
4	1.0	0.1	2.0	5.3±1.5^d
5	2.0	0.5	0.5	2.7±1.2^ef
6	0.5	1.0	1.0	7.3±1.5^c
7	2.0	0.1	1.0	3.3±1.5^e
8	0.5	0.5	2.0	6.0±1.7^cd
9	1.0	1.0	0.5	2.3±0.6^ef

Treatment	Plant growth regulators(mg·mL^-1)		Frequency of haustorium formation(%)	Number ofhaustorium
Treatment	2.4-D	KT	Frequency of haustorium formation(%)	Number ofhaustorium
1	0.00	0.25	0.0±0.0^b	0.0±0.0^c
2	0.25	0.25	33.3±23.1^b	3.7±0.6^b
3	0.50	0.25	66.7±30.6^a	5.3±2.5^ab
4	0.75	0.25	13.3±11.5^b	6.7±2.1^ab
5	1.00	0.25	16.7±5.8^b	8.3±1.5^a

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