Three new species of earthworms belonging to the genera Amynthas and Metaphire (Oligochaeta: Megascolecidae) from Yunnanı China Author Yuan, Zhu Author Dong, Yan Author Jiang, Jibao Author Qiu, Jiangping text Journal of Natural History 2019 2019-10-28 53 31 1961 1974 journal article 24130 10.1080/00222933.2019.1680760 959b13ea-8bec-4b77-b018-e50130773cf3 1464-5262 3654511 Metaphire daliensis Yuan & Dong sp. nov. Type material Holotype . One clitellate specimen ( YN201006-03 A): China , Yunnan , Dali city ( 25.94° N , 100.17° E ), 2010 m elevation, corn field by the stream, 10 August 2011 , Y. Yang. Other materials. Three clitellate specimens ( YN 201006 -03 B) with the same data as for holotype; three clitellate specimens ( YN 201101 -12 ): China , Yunnan , Yuanjiang National Nature Reserve ( 23.67° N , 101.85° E ), 854 m elevation, brown loam under orchard beside the stream, 16 July 2013 , J.B. Jiang, J. Sun, X.D. Lei and H.W . Feng ; eight clitellate specimens ( YN201103-01 ) and one clitellate specimen ( YN201103-04 ): China , Yunnan , Yuanjiang National Nature Reserve ( 23.55° N , 101.92° E ), 649 m elevation, black brown soil under the banana trees, 16 July 2013 , J.B. Jiang, J. Sun, X.D. Lei and H.W . Feng ; two clitellate specimens ( YN201107-05 ): China , Yunnan , Xishuangbanna Nature Reserve ( 21.61° N , 101.58° E ), 694 m elevation, red soil in tropical rain forest, 20 July 2013 , J.B. Jiang, J. Sun, X.D. Lei and H.W . Feng ; four clitellate specimens ( YN201624-02 ): China , Yunnan , Pu’ er ( 24.55° N , 100.53° E ), 1150 m asl , yellow soil, 29 July 2016 , X. Gao, Y.F. Lu, J.Z. Jiang ; one clitellate specimens ( YN201731-01 ): China , Yunnan , Ruili , Gaoligongshan Nature Reserve ( 26.57° N , 98.91° E ), 1429 m asl , 9 August 2017 , J.B. Jiang, Y. Dong, Q. Zhao, Z. Yuan ; one clitellate specimens ( YN201733-04 ): China , Yunnan , Ruili , Gaoligongshan Nature Reserve ( 25.71° N , 98.84° E ), 1279 m asl , 10 August 2017 , J.B. Jiang, Y. Dong, Q. Zhao, Z . Yuan ; two clitellate specimens ( YN201735-09 ): China , Yunnan , Ruili , Tongbiguan Nature Reserve ( 24.15° N , 98.03° E ), 957 m asl , 11 August 2017 , J.B. Jiang, Y. Dong, Q. Zhao, Z . Yuan ; four clitellate specimens ( YN201751-01 ): China , Yunnan , Ruili , Yongdedaxueshan Nature Reserve ( 24.03° N , 99.39° E ), 1125 m asl , 15 August 2017 , J.B. Jiang, Y. Dong, Q. Zhao, Z. Yuan . Figure 3. Metaphire daliensis Yuan & Dong sp. nov. (a) Spermathecal pore region; (b) Male pore region; (c) Spermathecae; (d) Intestine caecum; (e) Prostate gland. Table 3. A comparison of characters of Amynthas recavus sp. nov. , Amynthas yunlongensis (Chen & Xu, 1977) , and Amynthas exiguus (Gates, 1930) .

Character	A. recavus sp. nov.	A. yunlongensis	A. exiguus
Body length * width (mm)	58-64 * 2.1–2.3	58-102*4.5	24-48*2-3
Pigment
Dorsum	Pink before clitellum, light brown after clitellum	Brown flesh
Ventrum	No pigment	Brown flesh
First dorsal pore	12/13	12/13	12/13
Setal number in segment
Between male pores	9	?	9-15
Between spermathecal	12(VI), 12(VII), 12(VIII)	?	11-14(VI), 11–15(VII), 13–16(VIII)
pores
Spermathecal pores	Four pairs in 5/6-8/9, ventral, about 2/5 circumference	Four pairs in 5/6-8/9, about 2/5 circumference	Four pairs in 5/6-8/9, minute and superficial
ventrally apart from each other, eye-shaped	ventrally apart from each other, eye-shaped
Male pores	About 2/5 circumference ventrally apart from each	About 1/3 circumference ventrally apart from each	Minute and superficial, each pore at the centre
other, each on the top of a raised, elliptic, collapse-	other, each on a plain papilla, maybe one pair of	of a disc-shaped porophore that is slightly
topped porophore. One pair of oval, flat papillae in	small flat or raised papilla on XVII and XIX male	larger than the genital markings
presetal XVII,and three oval, flat papillae in presetal	pores line after setae ring
XIX.
Intestinal caeca	From XXVII to XXV, simple, finger-shaped, smooth	From XXVII to XXIII, simple	Simple,the margins smooth or with slight septal
ventrum	constrictions
Spermathecae	Ampulla thin heart-shaped, ampulla duct uniformly	Underdeveloped, only four pair granulation	Spermathecae with diverticular stalk nearly as
thick, the length about 4/5 of the ampulla. The	porophore	long as, to longer than the combined lengths
length of diverticulum about 3/5 of the main pouch	of duct and ampulla; seminal chamber small,
(duct and ampulla together), terminal 1/4 slightly	spheroidal to shortly ovoidal
dilated into an ovoid seminal chamber
Prostate glands	In 1/2XVII-XIX, relatively small, one single dense	Absent glandular part, only fine duct	?
racemose mass
Accessory glands near	Invisible	Invisible or underdeveloped or one block at XVII-	?
prostates	XIX

Figure 4. Neighbour-joining (NJ) tree of these species: based on combined analysis of COI and 12S sequences. Bootstrap values (1000 replicates) of NJ analyses are shown above/below the main lineages. Lumbricus rubellus was used as the outgroup taxa. Etymology The species is named after its type locality. Diagnosis Dimensions 110–143 mm by 3.9–7.2 mm ; segments 111–150. Setae numbering 28–40 at III, 30–44 at V, 46–64 at VIII, 58–74 at XX, 64–74 at XXV; 18–24 (XVIII) between male pores; 18–26 (VIII) between spermathecal pores. Spermathecal pores two pairs in 7/8-8/9, about 1/3 body circumference ventrally apart. Male pores in copulatory chambers at XVIII, opening of copulatory chambers about 1/3 body circumference ventrally apart, and a slightly extended papilla on the top, surrounded with 3–5 circular folds. Prostate glands located at XVII-XIX, underdeveloped. Spermathecae two pairs, in VIII and IX, about 3.3 mm . Ampulla heart-shaped, about 3.0 mm, ampulla duct short and thick. The length of diverticulum about 3.0 mm, terminal 1/2 slightly dilated into an elongated oval-shaped seminal chamber. External characters Dimensions 110–143 mm by 3.9–7.2 mm ; segments 111–150. No pigment anterior to segment VIII, ochre in posterior segments. First dorsal pore in 11/12. Prostomium 1/2 epilobous. Clitellum in segments XIV-XVI, beige, smooth and swollen, no setae. Setae numbering 28–40 at III, 30–44 at V, 46–64 at VIII, 58–74 at XX, 64–74 at XXV; 18–24 (XVIII) between male pores; 18–26 (VIII) between spermathecal pores. Setal formula: AA = 1.2–1.4 AB, ZZ = 1.6–2.0 ZY. Spermathecal pores two pairs in 7/8-8/9, about 1/3 circumference ventrally apart from each other. Female pore single in XIV. Male pores one pair in XVIII, about 1/3 circumference ventrally apart from each other, each located in the oval copulatory chamber, and a slightly extended papilla on the top, surrounded with 3–5 circular folds. In the copulatory chambers and male pores, two papillae on the left, one papilla on the right. Copulatory chambers with stalked glands. Internal characters Septa 5/6-7/8, thick and muscular, 10/11-12/13 slightly thickened, 8/9-9/10 absent. Gizzard bucket-shaped, in VIII-X. Intestine enlarged distinctly at XV. Intestine caeca start at XXVII, end in XXIII, between simple and complex, yellowish brown, smooth with two processes. Hearts four pair in X-XIII, the first pair relatively small, the others larger. The testis sacs minimum. Seminal vesicles two pairs, in XI-XII, the posterior pair is a little more developed than the anterior one. Prostate glands located at XVII-XIX, underdeveloped, ducts U-shaped, thicker at the distal part. Spermathecae two pairs, in VIII and IX, about 3.3 mm . Ampulla heart-shaped, about 3.0 mm, ampulla duct short and thick. The length of diverticulum about 3.0 mm, terminal 1/2 slightly dilated into an elongated oval-shaped seminal chamber. Remarks Metaphire daliensis sp. nov. belongs to the Metaphire insulana -group, in which there are only two species ( Metaphire insulana (Gates, 1930) and Metaphire leonoris (Chen, 1946)) according to Sims and Easton (1972) . A comparison of the three species ( Table 4 ), we observed that the spermathecal pores are two pairs, in 7/8-8/9, about 1/3-1/2 circumference apart ventrally; the shape of the ampulla is roughly the same with a short stalk. However, the first dorsal pore of M. daliensis is in 11/12, but M. insulana and M. lenoris are in 12/13; intestinal caeca of M. daliensis and M. insulana are between simple and complex, while M. leonoris is simple; the accessory glands near the prostate of M. daliensis and M. insulana are invisible, but M. leonoris is sessile in large masses. We also should compare the new species to the common species Metaphire californica (Kinberg, 1867) , which belongs to the Metaphire javanica -group. In the analysis of the K2P distances of combined COI and 12S sequences, the distance between M. daliensis and M. californica is 14.0% ( Table 5 ). However, the two species are in the same clade with 70% bootstrap support ( Figure 4 ). The two species have two pairs of spermathecal pores, in 7/8-8/9 and the first dorsal pores of both species are in 11/12. But, the most obvious differences between the species are the shape of the male pores and intestinal caeca. In the former, male pores are surrounded with three to five circular folds and the papillae in the interior of the copulatory chambers are small, and copulatory chambers are with stalked glands; the intestinal caeca are between simple and complicated. Male pores of M. californica are in simple lateral slits and copulatory chambers are without stalked glands; the intestinal caeca are simple, smooth on both edges.