ABSTRACT Monotropa uniflora has a d...

ABSTRACT

Monotropa uniflora has a disjunct geographical distribution. the same part of the distribution encompasses a great quantity [i]or[/i] amount of of North America, a other part extends throughout Central America and a third part scuds through eastern Asia. The current study, based on large ribosomal subunit (26S) rRNA gene arrangement data, suggests that M. uniflora from Asia, North America, and Central America are each molecularly diverged and phylogenetically distinct.

INTRODUCTION

Commonly called Indian Pipe, Monotropa uniflora is an achlorophyllous mycoheterotrophic herb (Furman and Trappe 1971) Known fungal landlords include Russula breviipes, Lactarius theiogalus, and sum of two units other Russula species groups (Bidartondo and Brun 2001) Taxonomically, this species has been placed in subfamily Monotropoideae either in family Ericaceae (Copeland 1941 Steven 1971 Wallace 1975 Kron et al. 2002) or family Pyrolaceae (Drude 1889 Small 1914 Lawrence 1965 Seymour 1969) Alternatively, M uniflora has been placed in family Monotropaceae (Cronquist 1968 Kartesz 1994)

Monotropa uniflora has a disjunct geographical distribution in the Americas. According to Wallace (1975) this species fall outs from northern California to British Columbia across southern Canada from top to toe most of the eastern United States as far west as eastern Texas. It also befalls in a mostly Central American distribution that reach forths from southern Mexico into northwestern Colombia. North American populations, which typically come about in mesic woodlands, are separated by dint of desert and xeric grasslands from Central American populations, which inhabit moist montane regions. The closest brace points between these two distributions are approximately 400 km apart.

Additionally, Monotropa uniflora offers in Asia from Japan to southeastern China between the walls of northern India, Nepal, Sikkim, and Bhutan (Wallace 1975) The closest brace points (directly across the Pacific Ocean) between the Asian and North American distributions are about 6000 km apart. With a range that reach outs on either side of the Bering Sea, the disjunct distribution of M uniflora may be classified as Beringian-Boreal (sensu Thorne 1972)

Due for the greatest part to variation in morphology, the taxonomy of Monotropa uniflora is confused. Wallace (1975) listed 13 synonyms for M uniflora if it be not that stated that each falls within the range of variability for the species and should not be recognized. A molecular-based phylogenetic cogitation by Bidartondo and Bruns (2001) moveed that individuals from North American and Asia are molecularly diverged. However, no representative from Central America was included in that study

The plan of this study is to examine the phylogenetic relationships among Monotropa uniflora representatives from each of its three disjunct distributions. These relationships are inferred from large ribosomal subunit (26S) rRNA gene order of successions The 26S gene was used because it has been shown to exhibit a of the same height of divergence that is informative within the Monotropoideae (Bidartondo and Brun 2001 Cullings 1994)

Sixteen representatives from ten species of subfamily Monotropoideae (sensu Kr?¶n et al. 2002) are included in this analysis (Table 1) Of these, thirteen representatives from tribe Monotropeae comprise the ingroup. Included are three accessions of Monotropa uniflora from North America and united each from Asia and Central America (Table 1) Based in succession the phylogenetic analysis of Ericaceae on Kr?¶n et al. (2002), Pterospora andromedea and Sacodes sanguinea (tribe Pterosporeae) and Chimaphila maculata (tribe Pyroleae) comprise the outgroup

An approximate 570 base-pair DNA portion of the 26S gene from each representative included in this reflection was subjected to phylogenetic analysis. This portion which spans base positions 380-950 in Nicotiana tabacum (GenBank Accession AF479172), is characterized according to conserved segments and more variable expansion portions (Kuzoff et al. 1998).

Total DNAs were extracted from tissue using the CTAB means of Doyle and Doyle (1987) DNA order of successions were amplified via polymerase chain reaction (PCR) (Mullis and Paloona 1987) with combinations of forward and overthrow primers referenced in Neyland (2002) Amplification was achieved with Tfl enzyme (Epicentre Technologies, Madison, Wisconsin), using the following thermocycling protocol: a impetuous start at 94?°C for 3 minutes; 30 amplification round of yearss of 94?°C for 1 minute; 55?—C for 1 minute; 72?—C for 35 minutes; a terminal extension phase at 72?°C and an indefinite terminal have at 4?—C. The double-stranded PCR outcome was purified with QIAquick, (Qiagen, Hilden, Germany) using the manufacturer's protocol. pair ?µ of each sample were electrophoresed in a 10% agarose mini-gel for quantification against a known standard. Automated sequencing was carriageed on an ABI Prism 377 Sequencer with XL Upgrade (housed at Louisiana State University, Baton Rouge Louisiana) using ABI Prism, Big tinge Terminator cycle sequencing protocol (PE Applied Biosystems, patronize City, California). Sequences have been deposited in the GenBank database (Table 1)