International Journal of Advanced Science and Technology Vol. 29, No. 7, (2020), pp. 4483-4493 Two new species of Beltrania, reported from forest flora of South Sagar forest division of Central India using morphological taxonomic techniques 1* 2* 3* 4* Atul Kumar Pandey , Ravindra Singh Thakur , Sandhya Parihar , Akhila Nand Rai *– Laboratory of mycology and plant pathology, Department of Botany, Dr. Harisingh Gour Vishwavidyalaya, Sagar, M.P. India E-mail of the Corresponding author – [email protected] Abstract: During the investigation of fungal flora in their terrestrial ecological niche, very conspicuous symptoms were observed on plants (Pyrus amygdalus Batsch, Mesua ferrea L.). After the microscopic study it was observed that both the plants were infected with the same fungal genus Beltrania. Later on these fungal organisms were studied in detail by using morpho – mycotaxonomic techniques and found that although they were the member of same genus but there morphological taxonomic attributes were different and on that basis they were supposed to be the new taxon of the species rank viz. Beltrania rosacearum sp. nov. and Beltrania calophyllacearum sp. nov. This paper provides the illustration and description of B. rosacearum sp. nov. and B. calophyllacearum sp. nov. Keyword: Beltrania rosacearum sp. nov., Beltrania calophyllacearum sp. nov., Pyrus amygdalus Batsch, Mesua ferrea L., morphological taxonomic attributes. Introduction: Beltraniaceae family was established by Nannizzi in 1934 to accommodate the single genus Beltrania but later on many worker add many genera such as Beltrania, Beltraniella, Beltraniomyces, Beltraniopsis, Parapleurotheciopsis, Porobeltraniella, Pseudobeltrania and Subramaniomyces (Maharachchikumbura et al. 2016), Hemibeltrania (Rajeshkumar et al. 2016), Subsessila (Lin et al. 2017) in family - Beltraniaceae. The genus Beltrania was established by O. Penzig in 1882. This is saprophytic genera, which members are collected from various habitats like dead leaves , other plant parts, air and soil. Very less members of this genus are known, because the members of this genera would like to grow under the leaf litter and collectors failed to notice them. Since they grow under the very closely spaced layers of litter then, very prominent setae are found in the members who play the role of space makers. The members of Beltrania genus has very conspicuous V- shaped or Kite Shaped conidia which are appendage while sometimes without appendage. In this paper we described two new species of Beltrania sp. reported from Sagar M.P. (Central India). Beltrania rosacearum sp. nov. and Beltrania calophyllacearum sp. nov on Pyrus amygdalus Batsch and Mesua ferrea L. respectively Materials and methods ISSN: 2005-4238 IJAST 4483 Copyright ⓒ 2020 SERSC International Journal of Advanced Science and Technology Vol. 29, No. 7, (2020), pp. 4483-4493 Fungal specimens were collected from the Botanic Garden of Sagar University, and brought to the laboratory in the zipped plastic bags to avoid cross contamination. processed by following standard techniques (Hawskworth 1974, Savile 1962). The surface scrapings of infected leaves were taken on glass slides and mounted in lactophenol cotton blue. The slides were observed in Leica Microscope (Leica DMLB). The Microphotographs of the interesting characters were photographed using Sony DSC W830 camera at different zooms for better picture quality with the help of different combination of eyepiece (5X, 10X, 15X) and objectives (4X & 40X) of Lieca Microscope. Some members of the Beltrania sp. were detailed morphologically studied by clicking photographs with Scanning Electron Microscope (SEM NOVA NANOSEM 450). For taking photographs with SEM specimens were dehydrated and coated with conducting material (Gold) to prevent charge build from Electron beam (Hall & Hawes 1991). The SEM microphotographs reveal the unseen structures of the fungus and their measurements which were not possible to be seen with the help of light compound microscope. Camera Lucida drawings were made and morphotaxonomic description was implemented as per current international standards. The taxonomic determinations were made with the help of relevant literature, and the description was compared with the earlier described allied forms so as to justify the novel entity of the collection. Results: Identification of Morphotypes: Taxonomic illustration and description Family – Beltraniaceae (Nannizzi, 1934) Genus – Beltrania (Penzig, 1882) (i) Beltrania rosacearum, Parihar, S., sp. nov Accession No. Holotype, HCIO No. 46,480 Etymology – Latin, Rosacearum named after the host family Rosaceae. Teleomorph: Undetermind. Anamorph: Hyphomycetous, Lesions hyphogenous, small to large, spreading over the whole leaf surface, brownish black. Colonies effuse, velutinous, brown to black, velvety. Mycelium immersed. Stroma usually present, erumpent often confined to epidermis. Setae, acute apex, simple dark brown smooth, thick walled, arising from the flat radially lobed basal cells, 165 × 4.5 – 7 µm. Conidiophores macronematous, mononematous, usually simple, straight to flexuous, pale olive to brown, smooth, septate, arising from the basal cell of the setae, 240 × 4.5 – 10 µm. Conidiogenous cell integrated, terminal, polyblastic, sympodial, clavate or cylindrical, denticulate, separating cells swollen. Conidia solitary, acropleurogenous, biconic, appendiculate, the free end being usually apiculate, aseptate, smooth, dark olivaceous to brown, with a hyaline transverse band, 29 – 16.5 × 16.5 – 5.0 µm. Appendage 13 – 6.5 µm long. Type: Collected from the dry leaves of Pyrus amygdalus Batsch., from the botanic garden of Sagar University, Sagar, M.P, India, leg. S. Parihar, S.U. Herb No. SRR – 400, Holotype, HCIO No. 46480. ISSN: 2005-4238 IJAST 4484 Copyright ⓒ 2020 SERSC International Journal of Advanced Science and Technology Vol. 29, No. 7, (2020), pp. 4483-4493 Figure 1. Camera lucida drawing showing A – Infected leaf, B – Setae attached with the basal cell, C – Setae with conidiophore (ii) Beltrania calophyllacearum Atul Kumar Pandey, A.N. Rai, R.S. Thakur, P. N. Singh and S.K.Singh, sp. nov. Accession No. - A.R.I. 9937, Mycobank Accession no. MB 827377 Etymology – Latin, specific epithet calophyllacearum derived from the host plant family. Teleomorph: Undetermind Anamorph: Leaf lesions amphigenous, predominantly hyphogenous, circular, irregular, starting from margin spreading towards the anterior, coalescing, light brown to black. Colonies hyphophyllous, represented by fine dots. Mycelium partly superficial, immerged in the host tissue. The hyaline hyphae ramify from the host tissue by forming plectenchymatous stromal structure. Setae and conidiophores both arise from flat swollen stromatal cells. Setae simple, 1–2 in number, erect, brown to black, smooth walled, 2–3 transversely septate, arising from basal lobed cells, base flattend, tapering towards length, tip acute to subobtuse, 55 × 117.5 µm, tip and base measure from 2.5 µm and 5.0 µm respectively. Conidiophores arising singly or in the groups of 2–3 from the basal stromatal cells, narrower towards base, apex swollen, macronematous, mononematous, simple, hyaline to olivaceous brown, 1–2 septate, denticulate, 17.5– ISSN: 2005-4238 IJAST 4485 Copyright ⓒ 2020 SERSC International Journal of Advanced Science and Technology Vol. 29, No. 7, (2020), pp. 4483-4493 25×2.5 –5 µm. Conidiogenous cells integrated, terminal, acropleurogenous, small, denticulate, subhyaline. Conidia rhomboidal smooth walled, acropleurogenous, appendiculate, appendage 5–7.5 µm long, hyaline to olive-brown with transverse banded septum, 17.5 – 27.5 ×7.5 –12.5 µm. Type: On healthy living leaves of Mesua ferrea L. (Calophyllaceae), Botanic Garden, Sagar University, South sagar forest division (M.P.) India, November 2017, (holotype, AMH-9937 isotype S.U. Herb No. Bot. PR 185). Figure 2. Microphotograph of Beltrania calophyllacearum sp. nov. AMH-9937 (holotype) a ̶ Habit of host plant, b- Symptom, c – Setae with obtuse apex (arrow) & conidiophore , d- Setae showing flat base & acute apex, e- conidiophores attach with conidia, f- conidia, coidiophores & setae with obtuse apex (arrow),g- conidiophores, h- Numerous conidia, Scale bar c-g = 20 µm. ISSN: 2005-4238 IJAST 4486 Copyright ⓒ 2020 SERSC International Journal of Advanced Science and Technology Vol. 29, No. 7, (2020), pp. 4483-4493 Figure 3. Beltrania calophyllacearum sp. nov. (Under SEM) a-d- Conidiophores, setae and conidia, e- conidiophore showing terminal dentate conidiogenous cells., f- Magnified view of conidia marked with length and width, g- Numerous appendaged conidia in low magnification, h- Conidia marked with length and width. ISSN: 2005-4238 IJAST 4487 Copyright ⓒ 2020 SERSC International Journal of Advanced Science and Technology Vol. 29, No. 7, (2020), pp. 4483-4493 Figure 4. Camera Lucida drawing of Beltrania calophyllacearum sp. nov .a: Appendaged conidia, b: Germinating conidia showing median lateral germ tube, c: Setae with conidiophores and appendaged conidia. Discussion – A thorough survey of available literature on genus Beltrania (M. B. Ellis 1971, Crous et al. 2014, Pirozynski 1969) has been done to show the distinct identity of the described species. A critical observation of the tabular data (Table: 1) comprising the important morphotaxonomic characters of different Beltrania species Beltrania rosacearum sp. nov. has shorter setae, much longer and broader conidiophores with broader conidia which make this distinct from