Figure 1.
Phylogenetic tree of single-cell SSU rRNA barcodes from Trichonympha isolated from the hindgut of Zootermopsis angusticollis.
Images are examples of manually isolated cells with arrows pointing to the SSU sequences obtained from these single cells. SSU rRNA sequences from Trichonympha species from Zootermopsis nevadensis were included for reference (T. sphaerica = AB434784, T. campanula = AB434812, and T. collaris = AB434783). AF023622 is from T. collaris isolated from the hindgut of Z. angusticollis. Also included are representatives of the next most closely related SSU rRNA sequences available: T. agilis from Reticulitermes speratus = AB003920, T. sp. from Hodotermopsis sjoestedti = AB326373, T. sp. from Reticulitermes santonensis = AB434787, T. tabogae from Incisitermes tabogae = AB434793. The best ML tree is shown. Numbers at nodes indicate ML bootstrap support and Bayesian posterior probability values. Statistical support is shown only for nodes with >70% bootstrap support and >0.90 posterior probability.
Figure 2.
Comparing SSU and ITS barcodes from manually isolated Trichonympha cells from Zootermopsis angusticollis.
A) Pairwise similarity matrix for DNA sequences from Trichonympha species. The upper and lower values are for SSU and ITS sequences, respectively. The mean pairwise similarities (± standard deviation) for comparisons of sequences within and between species are reported. B) Phylogenetic tree of single-cell ITS barcodes from Trichonympha isolated from the hindgut of Zootermopsis angusticollis. Sequences from Hexamastix mitis and Monocercomonas colubrorum were included to root the tree. The best ML tree is shown. Numbers at nodes indicate ML bootstrap support and Bayesian posterior probability values. Statistical support is shown only for nodes with >70% bootstrap support and >0.90 posterior probability. Statistical support for the T. campanula node (in italics) is also shown which does not satisfy the above criteria.
Figure 3.
Differential interference contrast (DIC) light micrographs of Trichonympha species in the hindgut of Zootermopsis angusticollis.
(A & B) Two cells matching the morphological description of T. postcylindrica sp. nov. These very large Trichonympha cells average 180 µm in length and are distinguished from other Trichonympha species in Z. angusticollis primarily by a posterior cylindrical projection (see detail in C). Also visible is a single posterior nucleus and a distinctly non-granular ectoplasmic region at the periphery of the anterior flagellated zone. (D–F) DIC micrographs for comparison of the three previously recognized species of Trichonympha in Z. angusticollis, (D) T. campanula (note the larger length:width ratio compared to T. collaris, posterior nucleus, and nondescript posterior end, (E) T. collaris (note the smaller length:width ratio and less posterior nucleus compared to T. campanula), and (F) T. sphaerica (note the smaller size, spherical cell shape, and anterior nucleus). All scale bars are 50 µm.
Figure 4.
Differential interference contrast (DIC) light micrographs of Trichomitopsis morphotypes in the hindgut of Zootermopsis angusticollis.
(A–C) Smaller morphotypes of Trichomitopsis (approximately 25 µm in diameter) corresponding to either T. parva or T. minor, which at present can only be distinguished from one another using molecular data. (D) Trichomitopsis termopsidis which is distinguishable by its much larger size. All scale bars are 20 µm.
Figure 5.
Phylogenetic tree of single-cell SSU rRNA barcodes from Trichomitopsis isolated from the hindgut of Zootermopsis angusticollis.
Sequences from Pseudotrypanosoma giganteum (AF052703 from Porotermes adamsoni) and Pentatrichomonas hominis (DQ412642 from the preputial cavity of the domestic cattle, Bos taurus) were included to root the tree. The best ML tree is shown. Numbers at nodes indicate ML bootstrap support and Bayesian posterior probability values. Statistical support is shown only for nodes with >70% bootstrap support and >0.90 posterior probability.
Figure 6.
Comparing SSU and ITS barcodes from manually isolated Trichomitopsis cells from Zootermopsis angusticollis.
A) Pairwise similarity matrix for DNA sequences from Trichomitopsis species. The upper and lower values are for SSU and ITS sequences, respectively. The mean pairwise similarities (± standard deviation) for comparisons of sequences within and between species are reported. B) Phylogenetic tree of single-cell ITS barcodes from Trichomitopsis isolated from the hindgut of Zootermopsis angusticollis. Sequences from Pentatrichomonas hominis and Trichomonas tenax were included to root the tree. The best ML tree is shown. Numbers at nodes indicate ML bootstrap support and Bayesian posterior probability values. Statistical support is shown only for nodes with >70% bootstrap support and >0.90 posterior probability.