Genetic variation of Macrobrachium lanchesteri (De Man, 1911) in Northeastern Thailand

Figures & data

Table 1. Collection localities of M. lanchesteri used in this study

Species	Location	Code	Geographic region	Latitude/longitude	Total (N)
M. lanchesteri	Mancha Khiri,	KK	Northeast	16º07ʹ28.5”N	115
	Khon Kaen			102º33ʹ39.0”E
	Thawat Buri,	RE	Northeast	16º02ʹ23”N	112
	Roi Et			103º44ʹ49”E
	Noen Sa-nga,	CP	Northeast	15º37ʹ18.6”N	112
	Chaiyaphum			102º04ʹ19.6”E
	Kantharawichai,	MS	Northeast	16º13ʹ56.6”N	110
	Maha Sarakham			103º16ʹ07.9”E
	Keang Sanam Nang,	NR	Northeast	15º38ʹ01.2”N	110
	Nakhon Ratchasima			102º11ʹ53.5”E
	Si Chiang Mai,	NK	Northeast	17º58ʹ08.7”N	110
	Nong Khai			102º27ʹ32.8”E
	Satuek,	BR	Northeast	15º18ʹ14.1”N	110
	Buri Ram			103º16ʹ57.8”E
	Na Wamg,	NB	Northeast	17º24ʹ32.4”N	110
	Nong Bua Lam Phu			102º04ʹ43.4”E
	Chiang Khan,	L	Northeast	17º54ʹ20.7”N	110
	Loei			101º42ʹ08.4”E
	Kanthararom,	SK	Northeast	15º08ʹ21.7”N	110
	Si Saket			104º31ʹ51.6”E
	Tha Tum,	SR	Northeast	15º15ʹ37.4”N	110
	Surin			103º33ʹ16.5”E
Total					1,219

Figure 1. Geographical location sites of M. lanchesteri in Northeastern Thailand.

Table 2. Water quality analyses from 11 locations in Northeastern Thailand

Location code	Water quality parameters
	Color	Temperature (ºC)	pH (pH)	Total hardness (mg/l)	Dissolved oxygen (mg/l)
KK	Clear	29	6	72	6.5
RE	Yellow wish	31	7	75	6.0
CP	Clear	28	7	67	5.0
MS	Clear	32	8	66	5.5
NR	Yellow wish	25	9	90	5.0
NK	Yellow wish	28	7	77	7.0
BR	Clear	31	6	88	6.5
NB	Clear	29	7	92	6.0
L	Clear	30	7	101	6.0
SK	Yellow wish	32	7	79	6.5
SR	light green	26	6	90	6.5
Mean	-	29.18	7	81.54	6.04

Table 3. Morphometric characters for male of M. lanchesteri 11 locations in Northeastern Thailand

Location	Morphometric character (cm)
	Total length		Standard length		Carapace length		Abdomen length		Telson length
	Mean±SD	range	Mean±SD	range	Mean±SD	range	Mean±SD	range	Mean±SD	range
CP	2.53 ± 0.12	2.3–2.7	1.95 ± 0.09	1.8–2.1	0.58 ± 0.06	0.5–0.7	1.35 ± 0.07	1.2–1.5	0.52 ± 0.06	0.2–0.4
NR	2.51 ± 0.12	2.3–2.7	1.93 ± 0.08	1.8–2.1	0.54 ± 0.12	0.4–0.7	1.39 ± 0.04	1.3–1.5	0.56 ± 0.06	0.5–0.7
KK	2.28 ± 0.25	2.5–3.4	2.14 ± 0.19	1.9–2.7	0.62 ± 0.12	0.4–1.0	1.49 ± 0.12	1.3–1.8	0.59 ± 0.14	0.4–1.0
MS	3.19 ± 0.22	2.7–3.7	2.38 ± 0.11	2.1–2.7	0.78 ± 0.08	0.7–1.0	1.56 ± 0.06	1.4–1.7	0.78 ± 0.12	0.6–1.0
RE	3.45 ± 0.13	3.2–3.7	2.49 ± 0.09	2.4–2.7	0.86 ± 0.06	0.8–1.0	1.62 ± 0.06	1.5–1.7	0.98 ± 0.06	0.8–1.1
NB	1.62 ± 0.12	1.5–1.9	1.50 ± 0.08	1.3–1.7	0.34 ± 0.12	0.3–0.4	1.03 ± 0.04	0.9–1.3	0.24 ± 0.06	0.2–0.3
BR	1.62 ± 0.08	1.4–1.8	1.46 ± 0.10	1.2–1.6	0.34 ± 0.05	0.3–0.4	1.13 ± 0.08	0.9–1.2	0.23 ± 0.04	0.2–0.3
SR	3.22 ± 0.22	2.9–3.6	2.38 ± 0.13	2.0–2.6	0.78 ± 0.08	0.7–0.9	1.63 ± 0.21	0.7–1.8	0.84 ± 0.12	0.7–1.0
L	3.43 ± 0.13	3.1–3.7	2.48 ± 0.09	2.4–2.7	0.85 ± 0.08	0.7–1.0	1.74 ± 0.07	1.6–1.9	0.95 ± 0.07	0.7–1.0
SK	2.45 ± 0.12	2.2–2.7	1.88 ± 0.06	1.8–2.0	0.50 ± 0.07	0.3–0.6	1.45 ± 0.05	1.3–1.5	0.55 ± 0.05	0.5–0.6
NK	3.86 ± 0.23	3.5–4.3	2.68 ± 0.17	2.3–3.0	0.98 ± 0.07	0.8–1.1	1.83 ± 0.12	1.7–2.0	1.08 ± 0.09	1.0–1.3
F-value	454.55*		284.00*		182.97*		134.32*		254.27*

* Significant at the 5% level at p≤ 0.05.

Table 4. Morphometric characters for female of M. lanchesteri 11 locations in Northeastern Thailand

Location	Morphometric character (cm)
	Total length		Standard length		Carapace length		Abdomen length		Telson length
	Mean±SD	range	Mean±SD	range	Mean±SD	range	Mean±SD	range	Mean±SD	range
CP	2.69 ± 0.12	2.5–2.9	2.01 ± 0.08	1.9–2.2	0.61 ± 0.08	0.5–0.7	1.45 ± 0.05	1.4–1.6	0.62 ± 0.05	0.5–0.7
NR	2.68 ± 0.10	2.5–2.9	2.03 ± 0.08	1.9–2.2	0.63 ± 0.07	0.5–0.7	1.40 ± 0.04	1.3–1.5	0.64 ± 0.05	0.6–0.7
KK	2.88 ± 0.30	2.4–3.5	2.19 ± 0.20	1.9–2.7	0.71 ± 0.14	0.5–1.0	1.51 ± 0.15	1.4–1.9	0.70 ± 0.14	0.6–1.1
MS	3.37 ± 0.18	3.0–3.7	2.42 ± 0.08	2.2–2.6	0.82 ± 0.09	0.7–1.0	1.60 ± 0.14	1.0–1.8	0.84 ± 0.10	0.7–1.0
RE	3.52 ± 0.12	3.3–3.8	2.56 ± 0.11	2.4–2.7	0.89 ± 0.09	0.7–1.0	1.75 ± 0.09	1.5–1.8	0.97 ± 0.07	0.8–1.1
NB	1.82 ± 0.10	1.7–2.1	1.61 ± 0.14	1.4–1.9	0.38 ± 0.04	0.3–0.4	1.08 ± 0.14	0.9–1.4	0.25 ± 0.05	0.2–0.3
BR	1.69 ± 0.10	1.5–1.8	1.55 ± 0.08	1.4–1.7	0.36 ± 0.04	0.3–0.4	1.16 ± 0.10	1.0–1.3	0.26 ± 0.04	0.2–0.3
SR	3.31 ± 0.34	2.1–3.8	2.48 ± 0.09	2.3–2.7	0.82 ± 0.09	0.7–1.0	1.66 ± 0.11	1.4–1.9	0.69 ± 0.19	0.7–1.1
L	3.50 ± 0.19	3.1–3.8	2.50 ± 0.10	2.4–2.7	0.89 ± 0.08	0.8–1.0	1.75 ± 0.08	1.6–1.9	0.97 ± 0.21	0.7–1.9
SK	2.55 ± 0.11	2.3–2.7	1.97 ± 0.08	1.8–2.1	0.51 ± 0.08	0.4–0.7	1.46 ± 0.07	1.4–1.6	0.54 ± 0.05	0.5–0.6
NK	3.90 ± 0.29	3.5–4.5	2.88 ± 0.21	2.6–3.5	1.02 ± 0.15	0.8–1.3	1.88 ± 0.12	1.7–2.0	1.14 ± 0.12	1.0–1.3
F-value	306.27*		145.51*		123.41*		122.60*		245.66*

*Significant at the 5% level at p≤ 0.05.

Table 5. Total length of M. lanchesteri 11 location in Northeastern Thailand

Location	(N)	Sex	Length range	Mean total	Variance
			(cm)	length (cm)	(cm)
CP	50	male	2.30–2.70	2.53 ± 0.12	0.016
	50	female	2.50–2.90	2.69 ± 0.12	0.015
NR	50	male	2.30–2.70	2.51 ± 0.12	0.014
	50	female	2.50–2.90	2.68 ± 0.10	0.012
KK	50	male	2.50–3.40	2.82 ± 0.25	0.063
	50	female	2.40–3.50	2.88 ± 0.30	0.094
MS	50	male	2.70–3.70	3.19 ± 0.22	0.051
	50	female	3.00–3.70	3.37 ± 0.18	0.033
RE	50	male	3.20–3.70	3.45 ± 0.13	0.018
	50	female	2.40–3.00	2.68 ± 0.15	0.023
BR	50	male	1.40–1.80	1.62 ± 0.08	0.008
	50	female	1.50–1.80	1.69 ± 0.10	0.011
SR	50	male	2.90–3.60	3.22 ± 0.22	0.051
	50	female	2.10–3.80	3.31 ± 0.34	0.12
L	50	male	3.10–3.70	3.43 ± 0.13	0.018
	50	female	3.10–3.80	3.50 ± 0.19	0.039
NK	50	male	3.50–4.30	3.86 ± 0.23	0.053
	50	female	3.50–4.50	3.90 ± 0.29	0.085
NB	50	male	1.50–1.90	1.62 ± 0.11	0.014
	50	female	1.70–2.10	1.82 ± 0.10	0.011
SK	50	male	2.20–2.70	2.45 ± 0.12	0.016
	50	female	2.30–2.70	2.55 ± 0.11	0.013

Figure 2. The 710 bp PCR product using COI primer of M. lanchesteri (100 bp DNA ladder, lane1–11 = M. lanchesteri, lane 12 = M. rosenbergii (out group)).

Figure 3. PCR-RFLP pattern of COI primer digested with Dde I(a), Alu I(b), Hinf I(c), Bgl II(d) and Hae III(e) (lane1 = uncut, lane2–10 = M. lanchesteri, lane11 = M. rosenbergii).

Table 6. Size of digested DNA fragments of M. lanchesteri and M. rosenbergii (out group) upon digestion with five restriction enzyme, Dde I, Alu I, Hinf I, Bgl II and Hae III

Enzyme	Pattern observed (bp)	M. lanchesteri	Out group
		(%)	(%)
Dde I	A: 450,350	+(100)	-
	B: 430,310	-	+(100)
Alu I	A: 400, 350,300	+(100)	-
	B: 710	-	+(100)
Hinf I	A: 300, 290, 150	+(100)	-
	B: 450, 290	-	+(100)
Bgl II	A: 620	+(100)	-
	B: 710	-	+(100)
Hae III	A: 490, 180	+(100)	-
	B: 350, 290	-	+(100)

*+, found in investigated species and in parenthesis represent percentages.

Table 7. Molecular taxonomic key base on composite haplotypes of COI gene of M. lanchesteri and M. rosenbergii (out group) digested with Dde I, Alu I, Hinf I, Bgl II and Hae III, respectively

Species	Composite haplotype
M. lanchesteri (CP, NR, KK, MS, RE,	AAAAA (100%)
NK, BR, SR, L, NB and SK)
M. rosenbergii	BBBBB (100%)

Table 8. Genetic distance of M. lanchesteri based on mitochondrial cytochrome c oxidase I (COI) gene sequence

location	BR	CP	NR	KK	MS	RE	L	NK	NB	SK	SR
BR
CP	0.035
NR	0.029	0.006
KK	0.033	0.001	0.004
MS	0.030	0.004	0.001	0.006
RE	0.033	0.001	0.004	0.000	0.006
L	0.017	0.018	0.012	0.016	0.013	0.016
NK	0.030	0.007	0.001	0.006	0.003	0.006	0.013
NB	0.010	0.024	0.018	0.023	0.020	0.023	0.006	0.020
SK	0.017	0.018	0.012	0.016	0.013	0.016	0.003	0.013	0.006
SR	0.013	0.024	0.018	0.023	0.020	0.023	0.006	0.016	0.006	0.006

Figure 4. Phylogenetic tree for M. lanchesteri in Northeastern Thailand based on unweighted pair group method using arithmetic average (UPGMA) analysis of COI gene sequences. Numbers are significant values for 1000 bootstraps.