Zoosyst. Evol. 100 (3) 2024, 913-922 | DOI 10.3897/zse.100.126340 eee BERLIN A new species of Grandinenia Minato & Chen, 1984 (Gastropoda, Stylommatophora, Clausiliidae, Garnieriinae) from Guangxi, China Zhong-Guang Chen", Jiao Jiang**, Ran-Xi Lin*, Guang-Long Xie*, Yu-Ting Dai‘, Xiao-Ping Wu’, Shan Ouyang? 1 School of Life Sciences, Nanchang University, Nanchang, Jiangxi 330031, China 2 Zhejiang Museum of Natural History, Hangzhou, Zhejiang 310012, China 3 State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangdong Laboratory for Lingnan Modern Agriculture, College of Agriculture, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China 4 School of Life Sciences, Qufu Normal University, Qufu, Shandong 273165, China https://zoobank. org/CA2E9204-BB04-4AF9-964C-910DCFAE7A8D Corresponding authors: Jiao Jiang (149152414@qq.com); Shan Ouyang (ouys1963@qq.com) Academic editor: Frank Kohler # Received 27 April 2024 # Accepted 20 June 2024 Published 8 July 2024 Abstract A molecular phylogenetic study was conducted on genus Grandinenia, based on COI and /6S sequences. A total of eight out of 26 Species in the genus, as well as three unidentified species were sequenced. Phylogenetic results supported the monophyly of Grand- inenia and the validity of all sampled species and subspecies. A new species, Grandinenia jiangjilini Chen, Lin, Wu & Ouyang, sp. nov., from Guangxi, southern China is identified and described, based on morphological comparison and molecular phylogeny. The discovery indicates that the inflated-fusiform shell is not isolated in genus Grandinenia and the species diversity of the genus still remains to be explored. Key Words Door snails, karst landscape, phylogeny, taxonomy Introduction The Guangxi Zhuang Autonomous Region is situated in southern China and is renowned for its distinctive land- scape and rich biodiversity. The karst landscape in this region provide a suitable habitat for land snails. The well-developed and exposed limestone have nurtured di- verse rock-dwelling gastropod groups, with genus Grand- inenia Minato & Chen, 1984 of subfamily Garnieriinae Boettger, 1926 being the most diverse and widespread in the region (Nordsieck 2012a, 2012b, 2012c, 2016; Lin and Lin 2022). * These authors contributed equally to this work. The subfamily Garnieriinae Boettger, 1926 1s a group of medium to large-sized door snails distributed from Myanmar to southern China. It is characterised by a fur- rowed neck, projected and unattached, so-called apos- trophic peristome and a lunella-type lunellar region (Nordsieck 2007). There are few molecular phylogenetic studies on the Garnieriinae and the limited studies suggest its close relationship with Synprophyminae Nordsieck, 2007 and Phaedusinae Wagner, 1922 (Uit de Weerd and Gittenberger 2013; Mamos et al. 2021; Uit de Weerd et al. 2023). Currently, Garnieriinae consist of seven gen- era, three of which are recorded in China: Garnieria Copyright Chen, Z.-G. et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 914 Bourguignat, 1877, Grandinenia Minato & Chen, 1984 and Tropidauchenia Lindholm, 1924 (Nordsieck 2012a, 2012b, 2012c). The genus Grandinenia is defined by the furrowed neck, the distinctly expanded and unattached peristome, the lunella-type lunellar and the inferior lamel- la separated from the superior lamella (Nordsieck 2007, 2012a). It is distributed in Laos, Vietnam and southern China and consists of 26 known species (Nordsieck 201 2a, 2012c, 2016; Grego et al. 2014; Lin and Lin 2022). Guanegx1 is the centre of diversity of the genus, with a total of 17 species recorded (Nordsieck 2012a, 2012c, 2016; Lin and Lin 2022). Most of these species were described in recent years, indicating that the species diversity has been poorly known in the past. Currently, the taxonomy of genus Grandinenia mainly rely on shell morphology, only two out of 26 species have been sequenced, but with- out detailed molecular phylogenetic studies. In this study, we conducted the molecular phylogenet- ic analysis of genus Grandinenia, based on partial COJ and /6S sequences and described a new species with a peculiar morphology from Guangxi, southern China. The discovery of this new taxon further increases the species diversity of land snails in Chinese karst landforms. Materials and methods Samples were collected from Guangxi and Guangdong, China from 2022 to 2023. Living specimens were initially frozen at -20 °C for 12 hours and subsequently thawed at room temperature for 12 hours to extract the soft parts. The soft parts were then fixed in 70% ethanol. Empty shells were cleaned, dried and preserved at 4 °C. All specimens were deposited in the School of Life Sciences, Nanchang University (Nanchang, Jiangxi, China). Photographs were taken by a Sony® Alpha a6500 Digital Camera and edit- ed in Adobe Photoshop CC 2015 (Adobe, San Jose, US). Maps were made in ArcGIS Pro (Esri, Redlands, US). Genomic DNA was extracted from foot tissues pre- served in 70% ethanol using a TIANamp Marine Animals DNA Kit (Tiangen Biotech, China). The quality and con- centration of the DNA were checked on 1% agarose gel electrophoresis and NanoDrop 2000 (Thermo Scientific, USA). Partial cytochrome c oxidase subunit 1 (CO/) and partial 16S ribosomal RNA (/6S) gene segments were amplified and sequenced for molecular phylogenetic analyses. Polymerase chain reaction (PCR) systems, con- ditions and primer pairs are listed in Table 1. Sequences were aligned using MEGA v. 6.0 (Tamura et al. 2013) and Chen, Z.-G. et al.: A new species of Grandinenia checked manually. The accession numbers of newly-ob- tained sequences and other species are given in Table 2. Phylogenies were reconstructed by the dataset com- bined CO/ and /6S genes using Maximum Likelihood (ML) and Bayesian Inference (BI). Five clausiliid spe- cles were used as outgroups for rooting the tree. ML analyses were performed in IQ-TREE v. 1.6.12 (Minh et al. 2013) using the Ultrafast fast bootstrap approach (Minh et al. 2013) with 10000 reiterations. The most appropriate model of sequence evolution (GTR+I+G for COI, GTR+G for /6S) was selected under Partiton- Finder2 v. 1.1 (Robert et al. 2017). Bayesian Inference (BI) was conducted in MrBayes v. 3.2.6 (Ronquist et al. 2012). The most appropriate model of sequence evolution (GTR+I+G) was selected under ModelFinder (Subha et al. 2017). Four simultaneous runs with four independent Markov Chain Monte Carlo (MCMC) were implemented for 10 million generations and trees were sampled every 1000 generations with a burn-in of 25%. The convergence was checked with the average standard deviation of split frequencies < 0.01 and the potential scale reduction factor (PSRF) ~ 1. Trees were visualised in FigTree v.1.4.3. Abbreviations NCU_XPWU Laboratory of Xiao-Ping Wu, Nanchang University (Nanchang, Jiangxi, China); cp clausilium plate; il inferior lamella; lu lunella; pp principal plica; se subcolumellar lamella; sl superior lamella; sp spiral lamella; At atrium; BC bursa copulatrix; BCD bursa copulatrix duct; D diverticulum; Ep epiphallus; FO free oviduct; P penis; PC penial caecum; PR penial retractor muscle; V vagina; VD vas deferens. Results Phylogenetic analyses A dataset consisting of 39 CO/ and 42 16S sequences from 11 species of Grandinenia, along with five outgroup taxa, was employed for phylogenetic analyses (Table 2). The aligned lengths of CO/ and /6S genes were 669 and 484 nucleotides. Within these sequences, 236 and 233 were revealed as vari- able sites, while 232 and 229 were designated as parsimony informative sites. Phylogenetic analyses generated ML and BI trees with congruent topologies (Fig. 1). Genus Grand- inenia forms a monophyly and further clustered into 12 dis- Table 1. Primer pairs and PCR conditions used in the analyses of the CO/ and 16S rRNA genes of Grandinenia. Genes Primer pairs Reaction systems Cycling conditions Reference CO! LCO1490: 12.5 ul 2 x Tag Plus Master Mix Il (Vazyme, Nanjing, China), 94°C: 2 min; 94°C: 10s,50°C: Folmer et al. GGTCAACAAATCATAAAGATATTGG 1 ul template DNA, 1 pl of each pair of primers, 9.5 plddH,O 60s, 72 °C: 1 min, 35 cycles; 72°C: — (1994) HCO2198: 10 min TAAACTTCAGGGTGACCAAAAAATCA 16S 16SA: CGGCCGCCTGTTTATCAAAAACAT =12.5 ul 2 x Taq Plus Master Mix Il (Vazyme, Nanjing, China), 94°C: 2 min;94°C:10s,50°C: — Pall-Gergely 16SB: GGAGCTCCGGTTTGAACTCAGATC 1 pl template DNA, 1 ul of each pair of primers, 9.5 pl ddH,O 60s, 72 °C: 1 min, 35 cycles; 72 °C: et al. (2019) zse.pensoft.net 10 min Zoosyst. Evol. 100 (3) 2024, 913-922 Table 2. GenBank accession numbers of the sequences for this study. Species Locality Col 16S References Grandinenia Lianggu, Qintang, PP473344 PP472576 This study rnthiga Guigang, Guangxl, p473345 PPA72577 This study China (type local- ity), 23°19'1'N, | PP473346 PP472578 This study 109°14'34°E = pP473347 PP472579 This study G. jiangyilini sp. Yao mountain, PP473375 PP472607 This study nov. Binyang, Nanning, 473376 PP472608 This study Guangxi, China, 23°26'12'N, PP473377 PP472609 This study 108°51'49'E = PP473378 PP472610 This study PP473379 PP472611 This study PP473380 PP472612 This study PP473381 PP472613 This study PP473382 PP472614 This study PP473383 PP472615 This study PP473384 PP472616 This study G. ookuboi Shanglin, Nan- PP473369 PP472601 This study pulchricosta ning, Guangxi, —_p473370 PPA72602 This study China, 23°27'9'N, 108°45'52"E PP473371 PP472603 This study G. rex Chenghuang, Xingye, PP473366 PP472598 This study Yulin, Guangxi, 473367 PP472599 This study China (type local- ity), 22°36'36'N, | PP473368 PP472600 This study 109°46'19"E G. cf. rutila Binyang, Nanning, PP473361 PP472593 This study ORIN PP473362 PP472594 This study 109°8'16"E G. fuchsi Guilin, Guangxi, PP473351 PP472583 This study China, 25°1835'N, p473352 PPA72584 This study 110°16'19°E PP473353 PP472585 This study tinct lineages. The phylogenetic relationships did not reflect a significant geographical correlation. Grandinenia mag- nilabris Nordsieck, 2012 from the middle northern Guangxi is the earliest diverging lineage. Grandinenia jiangjilini sp. nov. formed a distinct lineage, but its relationship within the genus was not well resolved (bootstrap supports = 55, pos- terior probabilities = 0.88). The genetic distances of COI se- quences between Grandinenia jiangjilini sp. nov. and other congeneric species ranged from 9.1% to 19.3% (Table 3). Species Locality col G. gastrum Lianggu, Qintang, PP473348 gastrum Guigang, Guangxi, PP473349 China (type locat_ ~—pp473350 ity), 23°18'51'N, 109°15'49"E G. gastrum Qintang, Guigang, PP473354 laticosta Guangxi, China, = PP473355 23°18°55'N, —— PP473356 109°16'30"E G. ignea Zhongshan, Hezhou, Guangxi, China (type locality), 24°27'48'N, LT O'35-E; G. magnilabris — Guzhai, Mashan, = PP473363 Nanning, Guangxi + PP473364 China (type loca = pp473365 ity), 23°41'7'N, 108°19'11"E G. sp. 1 Shanglin, Nanning, © PP473372 Guangxi, China, = PP473373 23°26.42'N, —— PPA73374 108°44'33'E G. sp. 2 Menggong, Qintang, PP473357 Guigang, Guangxi, PP473358 China, 23°10'52'N, 109°22'11"E Tropidauchenia —_ Huaiji, Zhaoqing, yanghaoi Guangxi, China (type locality), 23°55'18'N, 112°9'59"E T. orientalis Chongzuo, Guangxi, PP473359 China, 22°16'29"N, PP473360 107°4'14'E Agathylla goldi Europe KC756080 Alopia mariae Europe JQ911821 Isabellaria Europe AY425575 praestans Taxonomy Family Clausiliidae Gray, 1855 Subfamily Garnieriinae Boettger, 16S PP472580 PP472581 PP472582 PP472586 PP472587 PP472588 PP472617 PP472618 PP472619 PP472595 PP472596 PP472597 PP472604 PP472605 PP472606 PP472589 PP472590 PP472620 PP472621 PP472622 PP472591 PP472592 KF601271 1926 Genus Grandinenia Minato & Chen, 1984 Ako) References This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study Fehér et al. (2013b), Par- makelis et al. (2013) Fehér et al. (2013a) Uit de Weerd et al. (2004) Type species. Steatonenia mirifica Chen & Gao, 1982, by original designation. Table 3. Genetic distances of COI sequences computed by MEGA 6 of Grandinenia. 1 2 3 4 1 Grandinenia mirifica 0.006 2 G. jiangjilini sp. nov. 0.106 0.001 3 G. ookuboi pulchricosta 0.103 0.100 0 4 G. rex 0.097 0.091 0.087 0.001 5 G. cf. rutila 0.116 0.101 0.083 0.079 6 G. fuchsi 0.119 0.120 0.125 0.115 7 G. gastrum gastrum 0.055 0.119 0.121 0.102 8 G. gastrum laticosta 0.063 leas 0.131 0.119 9 G. magnilabris 0.198 0.193 CsA 0.183 10 G. sp. 1 0.097 0.100 0.016 0.083 11 G. sp. 2 0.118 0.142 0.130 0.115 5 6 7 8 9 0.001 0.131 0.003 0.132 0.122 0.003 0.135 0.124 0.036 0.004 0.207 0.216 0.207 0.209 0.005 0.088 0.124 0.118 0.128 0.206 0.130 0.145 0.127 0.145 0.200 10 11 0.001 0.130 0.001 zse.pensoft.net 916 90/0.96 55/0.88 76/0.93 73/0.91 80/0.99 93/0.99 75/0.92 100/0.98 97/0.97 62/0.93 100/0.98 Chen, Z.-G. et al.: A new species of Grandinenia Grandinenia sp. 1 Grandinenia sp. 1 Grandinenia sp. 1 - Grandinenia ookuboi pulchricosta Grandinenia ookuboi pulchricosta Grandinenia ookuboi pulchricosta , Grandinenia rex | Grandinenia rex Grandinenia rex eee Grandinenia cf. rutila = Grandinenia cf. rutila Grandinenia jiangjilini sp. nov. Grandinenia jiangjilini sp. nov. Grandinenia Jiangjilini sp. nov. Grandinenia jiangjilini sp. nov. Grandinenia jiangjilini sp. nov. Grandinenia jiangjilini sp. nov. Grandinenia jiangjilini sp. nov. Grandinenia siangjilini sp. nov. Grandinenia siangjilini sp. nov. Grandinenia jiangjilini sp. nov. Grandinenia fuchsi Grandinenia fuchsi Grandinenia fuchsi Grandinenia gastrum gastrum Grandinenia gastrum gastrum Grandinenia gastrum gastrum Grandinenia gastrum laticosta Grandinenia gastrum laticosta Grandinenia gastrum laticosta Grandinenia mirifica Grandinenia mirifica * Grandinenia mirifica Grandinenia mirifica Grandinenia ignea Grandinenia ignea Grandinenia ignea Grandinenia sp. 2 Grandinenia sp. 2 Grandinenia magnilabris Grandinenia magnilabris Grandinenia magnilabris Tropidauchenia yanghaoi Tropidauchenia yanghaoi Tropidauchenia yanghaoi Tropidauchenia orientalis Tropidauchenia orientalis Agathylla goldi Alopia mariae 100/1 100/1 99/1 lsabellaria praestans 0.05 Figure 1. Maximum Likelihood tree and Bayesian Inference tree inferred from CO/ and /6S gene sequences. Bootstrap supports/ posterior probabilities are shown on the left/right of nodes. Star shows the type species of the genus. Grandinenia jiangjilini Chen, Lin, Wu & Ouyang, sp. nov. https://zoobank. org/B234FD40-DDE4-4301-8724-E8FB819FCB81 Figs 2A, 3, 4A, B Type material. Holotype. 23 NCU_XPWU_YG01, Yao Mountain [®é1L1], Binyang County [eSH], Nanning City [Fd FT], Guangxi Zhuang Autonomous Region [ DPE BAK], China, 23°26'12"N, 108°51'49"E, leg. Zhong-Guang Chen, Ji-Lin Jiang & Guang-Long Xie, September 2023. Paratypes. 49 specimens, 23 NCU _XPWU_YG02-— 50, other information same as holotype. Different diagnosis. Shell entire (vs. decollated in G. ardouiniana (Heude, 1885), G. gabijakabi Grego & zse.pensoft.net Szekeres, 2014, G. gastrum (Nordsieck, 2005), G. mirifica (Chen & Gao, 1982), G. pallidissima Nordsieck, 2010, G. pseudofuchsi (Nordsieck, 2005), G. rex Nordsieck, 2007, G. rutila Nordsieck, 2016, G. schomburgi (Schmacker & Boettger, 1890), G. takagii (Chang, 2004), G. umbra (Chang, 2004)), hardly decollated, inflated-fusi- form (vs. slender-fusiform in all other congeners, except G. mirifica), light yellowish-brown, semitranslucent; te- leoconch with broad, blunt and sparse wrinkles (ribs) (vs. without or with thin and dense ribs in all other congeners); peristome not reflected; inferior lamella lower in front than within; penial caecum present (vs. absent in G. fuchsi (Gre- dler, 1883), G. pseudofuchsi, G. takagii and G. mirifica). Description. Shell (Figs 2A, 3A, B, 4A, B) (n = 50). Entire, with 8.75—9.5 whorls, hardly decollated, Zoosyst. Evol. 100 (3) 2024, 913-922 917 Figure 2. Grandinenia jiangjilini sp. nov. and two congeners. A. Grandinenia jiangjilini sp. nov., holotype (23, NCU_XPWU _ YGO1); B. G. mirifica, C. G. ignea. Figure 3. Detailed morphology of Grandinenia jiangjilini sp. nov. A—C. Shell morphology; D. Clausilium plate; E. Genital anat- omy. Abbreviations: cp clausilium plate; il inferior lamella; lu lunella; pp principal plica; sc subcolumellar lamella; sl superior lamella; sp spiral lamella; At atrium; BC bursa copulatrix; BCD bursa copulatrix duct; D diverticulum; Ep epiphallus; FO free oviduct; P penis; PC penial caecum; PR penial retractor muscle; V vagina; VD vas deferens. zse.pensoft.net O18 Chen, Z.-G. et al.: A new species of Grandinenia Figure 4. Living specimens of Grandinenia. A, B. inflated-fusiform, thin, fragile, semi-translucent, light yel- lowish-brown, with distinct darkish-red ribbon beneath the suture (fades quickly after fixing); dark seam along principal plica and lunella, body whorl in front of lunella darker; apical part conical to strongly attenuated. Suture deep. Protoconch smooth with 2.0—2.5 whorls. Wrinkles (ribs) on the teleoconch broad and blunt, most of them extending across the whole whorl, rather evenly distrib- uted and widely spaced; on the neck, riblets white, thin- ner, stronger, more widely spaced and undulate. Aperture vastly extended, oval. Peristome expanded, not reflected. Superior lamella continuous with spiral lamella without a curve. Inferior lamella visible in front view of the aper- ture, steeply ascending, moderately low to high within, it ends deeper than the end of spiral lamella. Subcolumellar lamella strong, bent, visible or not in front view of the ap- erture, ending less deeply than the end of inferior lamella. Lunella vertical, in oblique view, partly visible through the aperture. Principal plica short, initiates ventrolaterally and extending laterally, not reaching peristome. Clausili- um plate in oblique view nearly fully invisible, semi-trans- lucent; overall slender; stalk thin; plate relatively broad. Genitalia (Fig. 3C) (n= 10). Atrium short and relative- ly broad. Penis almost cylindrical and shortly narrower at transition to epiphallus. Penial caecum present. Epiphal- lus slender, shorter than penis and smaller diameter. Pe- nial retractor relatively thick and short, inserted at the middle part of penis. Vas deferens relatively slender and zse.pensoft.net randinenia jiangjilini sp. nov., C. G. gastrum; D. G. mirifica. short. Vagina thick, cylindrical, slightly longer than free oviduct. Basal part of diverticulum thick, rapid thinning to apical part and attached to spermoviduct. Spermovi- duct thick and long. Pedunculus of bursa copulatrix slen- der and long. Bursa copulatrix large, oval. Measurements. Holotype: shell height 24.5 mm, width 8.3 mm; aperture height 7.0 mm, width 7.7 mm. Paratypes: shell height 21.9—28.5 mm, width 7.4—8.4 mm; aperture height 5.9—7.3 mm, width 6.9-8.0 mm (n = 49). Etymology. The species is named after Mr Ji-Lin Ji- ang who first discovered the new species and assisted in the field survey. Vernacular name. {0 [C#} 2 (Pinyin: jiang shi xié guan luo). Distribution and ecology. Grandinenia jiangjilini sp. nov. is found from the Yao Mountain only (Figs 5-7). No other localities were found during the detailed sur- vey conducted in 2022—2023 of the surrounding hills. It inhabits the vertical limestone cliff together with Papil- liphaedusa porphyrea (Mollendorff, 1882) (Fig. 4A, B). Discussion The placement of the new species within Grandinenia is supported by both morphology (inferior lamella separat- ed from superior lamella) and molecular phylogeny. The absence of a comprehensive description of the genitalia, Zoosyst. Evol. 100 (3) 2024, 913-922 106°E 107°E 25°N 24°N 23°N 22°N 106°E 107°E 108°E o19 25°N 24°N 23°N 22°N 109°E 110°E 111°E Figure 5. Sample localities of Grandinenia used in this study. Star. Grandinenia jiangjilini sp. nov.; red point. G. mirifica, G. gas- trum gastrum and G. gastrum laticosta; blue point. G. ookuboi pulchricosta and G. sp. 1; green point. G. rex; purple point. G. cf. rutila;, yellow point. G. fuchsi; orange point. G. ignea; grey point. G. magnilabris; black point. G. sp. 2. shows the sampling locality. as well as the dearth of illustrations of lamellae and gen- italia in the most original descriptions of Grandinenia species, precludes the possibility of detailed comparison of the new species with most other congeners for these two characters. The comparison of the shell morphology of the new species with that of eight congeners collected in this study revealed that the lamellae of them are highly similar. In contrast to Tropidauchenia, no variation in the fusion or separation of lamellae was identified between 2S 1 1 } Boe Figure 6. Sampling locality. A. Karst hills surrounding the type locality; B. Type locality of Grandinenia Jiangjilini sp. nov. Arrow Grandinenia species. The new species is preliminarily dis- tinguished from G. fuchsi, G. pseudofuchsi, G. takagii and G. mirifica by the presence of penial caecum. However, the shell appearance of the new species is sufficiently dis- tinctive that it can be readily distinguished from all other congeners through a simple comparison. Grandinenia Ji- angjilini sp. nov. can be easily distinguished from all other congeners by the teleoconch with broad, blunt and sparse wrinkles (ribs) (vs. without or with thin and dense ribs). zse.pensoft.net 920 led Furthermore, except for G. mirifica, the remaining 25 species of Grandinenia exhibit a relatively slender shell (Fig. 2C). Grandinenia jiangjilini sp. nov. can be easily distinguished from them by the different shell shape (in- flated-fusiform vs. slender-fusiform). Grandinenia jiang- Jilini sp. nov. is most similar to G. mirifica by similar in- flated-fusiform shell (Fig. 2A, B), but differs by the broad and sparse wrinkles (ribs) on teleoconch (vs. thin and dense), peristome not reflected (vs. reflected), protoconch preserved (vs. decollated), shell semi-translucent and fragile (vs. opaque and solid) and the different shell co- lour (yellowish-brown vs. yellowish-white). Grandinenia jiangjilini sp. nov. is also somewhat similar to G. dautzen- bergi (Morlet, 1892) and G. yulinensis Nordsieck, 2012, but differs by the more inflated shell, the stronger ribs on neck, the deeper suture, the thinner-walled and more frag- ile shell and the broad, blunt and sparse wrinkles (ribs) on teleoconch (vs. with very weak ribs to even smooth). The validity of Grandinenia jiangjilini sp. nov. was also supported by the molecular phylogeny. It forms a distinct lineage and has a distant relationship with G. mir- ifica. The molecular phylogenetic relationships of genus Grandinenia do not correspond to the morphological sim- ilarities. The important characters of shell including shell shape, integrity, ribs, thickness and spiral ribbon, have homoplasiously evolved more than once. The species like G. rex, G. fuchsi and G. ignea with clearly similar smooth, thin, fragile and semi-translucent shells with spi- zse.pensoft.net Chen, Z.-G. et al.: A new species of Grandinenia ral ribbons, do not form a monophyletic group. The same phenomenon also occurs in species with similar thick and ribbed shells. The result shows that the shell appearance of Grandinenia is an effective means of distinguishing species, but does not reflect the interspecies affinities. The phylogenetic relationship also did not demonstrate a clear geographical correlation overall. Grandinenia mag- nilabris from the middle northern Guangxi (the western- most distribution of the sampled species) is the earliest diverging lineage. Grandinenia ignea from north-eastern Guangxi sistered with G. sp. 2 from the central region. In addition, G. fuchsi from north-eastern Guangxi was sis- tered with the clade, which consists of four species from the central region and one from the south-eastern region. The distribution pattern may be attributed to multiple in- dependent diffusions from west to east in history. Only two lineages exhibited a certain geographical correlation, G. sp. 1 and G. ookuboi pulchricosta from Shanglin and G. mirifica, G. gastrum gastrum and G. gastrum laticos- ta from Guigang, which formed monophyletic lineages, respectively. Due to the limited species included in this study, extensive and continuous sampling in future stud- ies may help further analysis of the phylogeny and eluci- dation of the reasons for its distribution pattern formation. The variation of shell morphology is frequently se- lected by environmental factors (Chiba 2004; Rolan-Al- varez 2007; Giokas et al. 2014). The shell characters of land snails play a pivotal role in regulating the water and Zoosyst. Evol. 100 (3) 2024, 913-922 heat budget, thereby preventing desiccation (Cowie and Jones 1985; Pfenninger et al. 2005; Giokas et al. 2014). The discovery of the new species shows that the special inflated-fusiform shells are not isolated in genus Grand- inenia. Phylogenetic result indicates that the new species and G. mirifica do not form a monophyletic group, sug- gesting that the similar shell shape between them may be the result of convergent evolution. Nordsieck (2012a) proposed that the inflated-fusiform shell of G. mirifica is an adaptation to a special habitat, but did not speci- fy what this habitat is. For Grandinenia, it is probable that the inflated-fusiform shells have increased resistance to desiccation and ultraviolet radiation compared to the slender-fusiform shells, although the precise mechanism of action remains unclear. Through field observation, it was found that the two shell shapes of Grandinenia corre- spond to two life strategies. The majority of Grandinenia Species with slender-fusiform shells burrow into crevices to hibernate during the dry season (Fig. 4C). It is chal- lenging to find them during the dry season, but a consid- erable number of individuals can be observed during the rainy season in the same area. In contrast, the two species with inflated-fusiform shells primarily hibernate on rock surfaces which can be found throughout the year, regard- less of precipitation patterns (Fig. 4A, B, D). The discovery of new taxon or just new species indi- cates that the species diversity of Grandinenia in Guangxi still remains to be explored. Nine species of Grandine- nia have been recoded within a few dozen kilometres of the new species’ type locality (Nordsieck 2012a, 2012c, 2016; Lin and Lin 2022) and several specimens which may represent other undescribed species were found during the field survey. Extensive exploration of land snails in Guangxi should be strengthened in the future which may lead to the discovery of yet-to-be-described species. In addition, the protection of Grandinenia should be a priority. As a rock-dwelling land snail, the survival of Grandinenia depends on the exposed rock environ- ment under the forest. Large-scale mining of limestone and agricultural reclamation in Guangxi pose a threat to it. It was even found that some peaks that used to be rich in Grandinenia have been completely blasted and disap- peared. The field survey conducted in 2024 revealed that the native shrubs in the type locality of Grandinenia ji- angjilini sp. nov. had been completely cut down (Fig. 7). This has resulted in the environment of the rock walls becoming more exposed and dry and has led to a signif- icant reduction in the population size of Grandinenia ji- angjilini sp. nov. It is imperative that further protection measures are implemented without delay, otherwise a sig- nificant number of unique species may be lost. 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