BIOGEOGRAPHY OF RUST FUNGI AND THEIR HOSTS IN PAKISTAN.
ABSTRACT: During the exploration of rust fungi in 2014, some of the hot spots of rust fungi, i.e. Swat district, Murree and Azad Jammu and Kashmir were visited, which resulted the identification of four genera of rust fungi naming Puccinia, Cerotelium, Miyagia and Uredo containing seven species on eight host plants. Among eight hosts, Artemisia vulgaris hosting Puccinia tanaceti is a new host record from Pakistan. This study will be useful for the future exploration of rust fungi across the Pakistan.
Key words: Khyber Pakhtunkhwa, Murree, AJK, Rust Fungi and Hosts, Pucciniales
Rust fungi (Basidiomycota) constitutes one of the major obligate biotrophic parasitic group of fungi in the order Pucciniales (formally Uredinales). They infect almost all types of plants either wild or cultivated with rusty symptoms on leaves, stems and fruits . The exploration of rust fungi in different parts of Pakistan is in progress and recently Swat district was studied by Pakistani mycologists who described about 65 rust species from Pakistan [2-3]. However, other hot spots of rust species such as Azad Jammu and Kashmir and Murree were studied long time ago, and still these regions are need to be explored more . The biodiversity of rust fungi is being increased due to genetic recombination and new strains are being identified throughout the Pakistan. In 2014, many field surveys were organized in Swat district, Murree and Azad Jammu and Kashmir for the exploration of rust fungi and their associated hosts.
As a result, eight host plants were reported for the first time from these regions i.e. six from Swat district and one from each Murree and Azad Jammu and Kashmir.
MATERIAL AND METHODS
Different regions of Pakistan were surveyed for the exploration of rust fungi during 2014. Infected plants were preserved and submitted to the herbarium of Plant Sciences Department, Quaid-i-Azam University, Islamabad, Pakistan (ISL), and rust spores were mounted in glycerin jelly and fixed as semi-permanent slides. The spores were measured and photographed at 100x and 1000x magnifications under a Leitz HM-LU compound light microscope. At least 30 spores were measured for each spore stage, including the smallest and the largest spores found. Host plants were identified by comparing new collections with botanical specimens held in herb. ISL. For Scanning Electron Microscopy (SEM) the spores were scraped from the leaves, mounted on a stub, coated with gold in a sputter-coater and examined with a JSM-5910 SEM.
RESULTS AND DISCUSSION
Puccinia menthae Pers., Syn. Meth. Fung. (Gottingen) 1: 227 (1801) SPERMOGONIA and AECIA not found. Uredinia abaxial, brown, rounded, scattered, 0.8-2 x 0.6-1 diameter. UREDINIOSPORES globose to sub-globose, obovoid or ellipsoid or elongated ellipsoid, hyaline to light brown, 20-32.5 x 15-22.5 um, wall 2-2.5 um thick, brown, germ tube hyaline, 4-7 x 3-4.5 um. Telia generally abaxial, rarely amphigenous, on leaves and petioles, globose, blackish brown, scattered, pulverulent, 0.5-1.6 x 0.4-0.6 um. TELIOSPORES 2-celled, ellipsoid to obovoid, spherical at both ends, slightly or not constricted at the septum, 22.5-27.5 x 20-22.5 um, chestnut brown, verrucose, wall 2-4.5um, thick at sides, papilla hyaline, 4-5 x 10-12.5 um, apex rounded, germ pores 2 and equatorial; Pedicel hyaline, 10-47.5 x 5-10 um, delicate.
On Origanum vulgare L. with II + III stages Pakistan, Khyber Pakhtunkhwa, Swat District, Alamganj, at 1500 m. a. s. 1, 15th November, 2014. BA# 01. On Mentha longifolia L. with II stage, Pakistan, Khyber Pakhtunkhwa, Swat District, Alamganj, at 1200 m. a. s. 1, 20th November, 2014 BA# 02.
Puccinia menthae has been reported on Origanum vulgare L. from Kaghan valley, Naran, Sharhan and Changa Gali; on Mentha sylvestris L. from Quetta, Chillian wala, Mingora, Poonch, Kaghan valley, Naran, Muree hills and Peshawar, on Calamintha umbrosa Rchb. and Calamintha clinopodium Benth. from Kaghan, Batakundi, Changla Gali, Swat, Kalam and Naran; on Neptea spp. from Kaghan valley [5-6], on Mentha longifolia and Nepeta campestris from Azad Jammu and Kashmir, Neelum valley and Sharda , on Neptea sp. from Swat .
Puccinia menthae is a new record on Mentha longifolia from Khyber Pakhtunkhwa, Pakistan. While on Origanum vulgare it is recorded for the first time from Swat district, KP, Pakistan.
Puccinia tanaceti DC., FI. Fr. 2, 222. (1805) SPERMOGONIA and AECIA not found. UREDINIA amphigenous, mostly hypophyllous, cinnamon brown, and scattered, rounded, 0.3-0.7 mm diameter, pulverulent. UREDINIOSPORES globose to ellipsoid, light brown, wall chestnut brown, 20-32.5 x 22.5-32.5 um, wall 1.5-2.5 um thick, echinulate, germ pores-3. TELIA also hypophyllous, blackish-brown; scattered or grouped, rounded or elliptic, 0.3-1 mm., pulverulent; TELIOSPORES ellipsoid or clavate, 40-60 x 22.5-27.5 um, circular at both ends or somewhat pointed above and tapered below, septa generally constricted, light brown, 2-celled, papilla brown, 5-10 x 15-22.5 um, wall thick, brown, 2-3.5 um, thick at the sides, upper cell pore is apical, lower cell pore is near the septum; pedicel elongated, persistent, hyaline, 17.5-125 x 12.5 um in size.
Material examined On Artemisia vulgaris L. with II and III stages, Pakistan, Khyber Pakhtunkhwa, Swat District, Charbagh, Alamganj at 1134 m. a. s. 1, 10th December, 2014. BA# 03.
Puccinia tanaceti on Artemisia spp. has been reported from Pakistan [8-9].
Puccinia tanaceti on Artesmisia vulgaris here reported for the first time in Pakistan
Uredo otostegiae S. Ahmad, Biologia Lahore 2 (1): 92 (1956) SPERMOGONIA, AECIA and TELIA not found. UREDINIA mainly hypophyllous with spots surrounded by light green rings, mostly in groups, confluent, rounded, dark brown, erumpent. UREDINIOSPORES are single celled, globose-ovoid or subglobose, 20-27.5 x 20-30 um, wall 1-1.5 um thick, light brown to reddish brown, smooth, germ pores equatorial and 1-2.
Material examined On Otostegia limbata (Benth.) Boiss., with II stage, Pakistan, Khyber Pakhtunkhwa, Swat District, Charbagh, Alamganj, at 1134 m. a. s. 1, 12th March, 2015. BA# 04. On Otostegia limbata, with II stage, Azad Jammu and Kashmir, Kotli, Poonch, at 1220 m. a. s. 1, 25th July, 2015. BA# 05.
Uredo otostegiae on Otostegia limbata has been reported from Poonch and Balakot [5-6]  , Katas and Salt range regions . According to literature  this rust species was first believed to have 2 germ pores, however, Abbasi  described 2-5 germ pores. In our findings, specimens from Khyber Pakhtunkhwa had 2 germ pores while specimens from Poonch (AJK) had 1 germ pore.
Otostegia limbata is reported as a host of Uredo otostegiae for the first time from Khyber Pakhtunkhwa, Pakistan.
Puccinia wattiana Barclay, Journal of the Asiatic Society of Bengal 54: 109 (1890)
SPERMOGONIA, AECIA and UREDINIA not found. Telia amphigenous, blackish, rounded, pulverulent, rarely in groups, naked, sometimes surrounded by ruptured epidermis. TELIOSPORES 2-celled abundant, sometimes 1-celled, dimorphic, ellipsoid or oblong, golden brown to blackish brown or chestnut, 20-27.5 (-35) x 17.5-20 um (24.4 x 17.7); wall chestnut brown or paler, smooth, 2-3 um thick, not constricted at the septum, spherical at both ends, germ pores 2, upper cell possess apical and lower cell possess near or adjacent to pedicel; papilla hyaline or light brown, 4-6.5 x 6-10 um, pedicel hyaline, deciduous, 10-98.5 x 6.5-9.5 um.
Material examined On Clematis grata Wall. with III stage, Pakistan, Khyber Pakhtunkhwa, District Swat, Madyan, at 1367 m. a. s. I, 04 October, 2014. BA# 06.
Puccinia wattiana has been reported on Clematis grata in Miana, Margazar (Swat, K.P.K) and Murree [5-6] on C. barballata Edgew., from Kawai (Kaghan Valley) [13-14], Leepa Valley, Azad Jammu and Kashmir .
Puccinia wattiana on Clematis grata is described from Swat District, KP, Pakistan.
Miyagia pseudosphaeria (Mont.) Jorst., Nytt Magasin for Botanik 9: 78 (1962)
SPERMOGONIA, AECIA and TELIA not found. UREDINIA mostly on leaves, sometimes on petioles and stems, amphiphylous, chiefly hypophylous, yellowish or pale yellowish brown, scattered or combined in groups, naked, round, about 1-2 cm in diam., subepidermal, Pulverulent; UREDINIOSPORES occurs in many shapes as spheroidal, oval, pyriform, sometimes sub-globular or irregular shapes and mostly with pigmented yellowish contents, some spores observed with flattened bases, (21-) 22.5-35 (-37) x (17-) 17.5-20 (-23) um (av. 28.5 x 18.66); walls thickened, hyaline, well echinulate and 2.5-4 um (4 um); PARAPHYSES were marginal, yellowish brown to reddish brown, cylindrical or clavate, 50-130 (-135) x 6-8 um thick, apically rounded; wall 1-3 um thick, side walls seems thicker at apex, hyaline below and sometimes dichotomously branched. Under Scanning electron Microscope the outer spore surface is completely echinulate (spines in prickles shape) and not aggregated or at some distance from each other.
Material examined On Sonchus arvensis L. with II stage, Pakistan, Punjab, Murree, Bunsra Gali, at 1777 m. a. s.1, 12 November, 2014. BA# 07.
Miyagia pseudosphaeria has been reported on Sonchus arvensis from Pakistan [8-10].
Miyagia pseudosphaeria is a new record from Murree, Pakistan.
Puccinia exhausta Dietel, Bot. Jb. 28: 283. (1900) SPERMOGONIA, AECIA and UREDINIA not found. Telia amphiphyllous, small, rounded, scattered or in circular groups, dark brown to blackish, erumpent or covered by epidermis. TELIOSPORES oblong or ellipsoid, conical or rounded at apex, basal mostly roundish, slightly constricted or not constricted at the septum, 37.5-47.5 x 22.5-25 um, sometimes 1-celled and elongated, usually two-celled, walls smooth, hyaline to yellowish and chestnut brown, spore wall thickness at the sides 2.0-3.0 um, apical wall may be thickened up to 3 um, 2-germ pores, germ pore of the upper cell is apical, of the lower cell nearly basal or adjacent to pedicel, with distinct hyaline papilla over the pore, 5-7.5 x 9-12 um, pedicel hyaline, fragile, 47-107 x 4-8.5 um.
Material examined On Clematis grata Wall. with III stage, Pakistan, Azad Jammu and Kashmir (AJK), Rawalakot, at 1637 m. a. s. 1. 5th December, 2014. BA# 08.
Puccinia exhausta was reported previously from different Clematis Spp. [15,16,17]. From Pakistan, it has previously been reported on Clematis Montana Buch. Ham. ex DC. from the Kawai and Malakandi forests (Kaghan valley) [13,8] and on Clematis grata from District Mansehra .
Puccinia exhausta on Clematis grata for the first time reported from Azad Jammu and Kashmir (AJK).
Cerotelium fici (Castagne) Arthur, Bull. Torrey Bot. Club 44: 509. 1917.
SPERMOGONIA, AECIA and TELIA not found. UREDINIA abaxial, distributed over the whole surface, small, occasionally in large irregular groups, pulverulent, slightly erumpent, reddish brown in color. UREDINIOSPORES are subglobular or broad-spheroidal to obovate or elliptical, yellowish brown, densely echinulate, 20-37.5 x 12.5-25 um, wall 1-2 um thick, germ pores found in scattered form, obscure.
Material examined On Ficus palmata Forssk., with III stage, Pakistan, Khyber Pakhtunkhwa, Swat district, Alamganj, at 700 m. a. s. 1, 25th February, 2015. BA# 09.
Cerotelium fici has been reported previously on Ficus carica, F. palmata, and F. religiosa L. from Lahore, Change Manga, Sangla Hill, Tandojam, Malir (Karachi), Rawalpindi and Mansehra [5-6,20,21,22,19], on Broussonetia papyrifera .
Cerotelium fici on Ficus palmata is a new record from Swat District, KP, Pakistan.
Pakistan has been gifted with rich flora in which rust fungi makes a significant portion. The research work on rust fungi is in progress based both on morphology and molecular level. However, Pakistan in terms of rust fungi, is still under exploration and the present study provides a status of the distribution of rust species on various hosts across Pakistan. In future it is vital to consider multi approach assessment for the precise and accurate study of rust fungi.
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