Molecular Diversity of Walnut (Juglans regia L.) Among Two Major Areas in Morocco in Contrast with Foreign Varieties

Figures & data

Table 1. List of walnut local clones and Bulgarian varieties included in this study

Genotype codes	Geographical origin	Locality, village name	Genotype codes	Geographical origin	Locality, Village name
NMA1			NMDT1
NMA3			NMDT2
NMA4			NMDT3
NAM1		Amizmiz Ait Ihla	NMDT4
NSF2		Sidi Fares	NMDT6
NIM1		Imlil	NMDT8
NIM2		Imlil	NMDT10
NAK		Akrsouak, Amghras	NMT1
NAO1		Ait Lkak, Oukaymiden
NAO2		Ait Lkak, Oukaymiden	Bulgarian genotypes codes	Variety names*
NAO3		Ait Lkak, Oukaymiden	DJ7	Djnovski
NAO4		Ait Lkak, Oukaymiden	IS1	Isvor
NAO5		Ait Lkak, Oukaymiden	SH1	Sheinovo
NI1		Imi N’Isli	SH7	Sheinovo
NI2		Imi N’Isli	DR7	Drianoski
NI3		Imi N’Isli	DR8	Drianoski
NIZ1		Imzyane	SI5	Silistrenski
NIZ2		Imzyane	SI8	Silistrenski
NIJ1		Ijoukak
NIJ5		Ijoukak
NIJ6		Ijoukak
NIJ7		Ijoukak
NOR2		Ourika
NOR3		Ourika
NOR5		Setti Fatma

*Among Bulgarian cultivars, some were introduced with similar denominations, however with different morphological profiles. Therefore, each one were given a different code in order to assess their molecular similarity.

Table 2. List of ISSR primers used and their annealing temperature

Primer	Sequence (5’-3’)	Annealing temperature (°C)
1MA5-3	(CA)8TG	52
F2	(CA)6GC	46
F3	(CA)6AG	46
F5	(AC)8CA	52
F6	(AC)8CG	56
F8	(AG)8CC	46
F13	(GA)8CG	52
F14	(GA)8CT	52
F15	(GT)8CC	46
IMA12	(CA)8TG	46
UBC808-2	(AG)8C	56
UBC818X	(CA)9G	52
UBC810	(GA)8T	52

Table 3. ISSR primers, optimal annealing temperatures, and amplification results of ISSR primers in 41 walnut genotypes

Primer	Séquence (5’-3’)	TR	AT (°C)	TB	NTB	NPB	NMB	EMR	MI	PPB	PIC
1MA5-3	(CA)8TG	Di	52	1305–206	10	10	0	10	2.72	100	0.272
F2	(CA)6GC	Di	46	765–197	7	4	3	2.29	1.336	57.14	0.334
F3	(CA)6AG	Di	46	1131–230	9	7	2	5.44	1.897	77.78	0.271
F5	(AC)8CA	Di	52	2834–567	10	7	3	4.90	2.59	70	0.37
F6	(AC)8CG	Di	56	2433–473	10	7	3	4.90	2.31	70	0.33
F8	(AG)8CC	Di	46	2446–367	8	8	0	8	2.792	100	0.349
F13	(GA)8CG	Di	52	1587–268	7	5	2	3.57	1.475	71.43	0.295
F14	(GA)8CT	Di	52	1944–180	8	6	2	4.50	1.272	75	0.212
F15	(GT)8CC	Di	46	2144–188	9	6	3	4	2.13	66.67	0.355
IMA12	(CA)8TG	Di	46	2160–208	13	9	4	6.23	3.114	69.23	0.346
UBC808-2	(AG)8C	Di	56	1837–167	10	6	4	3.60	1.944	60	0.324
UBC818X	(CA)9G	Di	52	2290–314	7	6	1	5.14	2.046	85.71	0.341
UBC810	(GA)8T	Di	52	2090–178	12	9	3	6.75	2.664	75	0.296
Mean					9.23	6.92	2.31	7.33	2.18	78.57	0.32

AT: Annealing temperatures; NPB: Number of polymorphic bands; PPB: Percentage of polymorphic bands; NTB; Nombre total of bandes; NMB: Number of monomorphic bands; TB: Taille of bandes; EMR: Effective multiple ratio; MI: Marker index; PIC: polymorphic information content.

Figure 1. Distribution of genetic similarity frequencies for all pairs among the 41walnut genotypes

Table 4. Heatmap of distances matrix between the 41 genotypes of walnut

Figure 2. Dendrogram screening genetic relationships between walnut genotypes based on ISSR markers