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Staub: W6743, W6744, W6745

Germplasm Release

Published in HortScience 31:1243-1245:1996

Cucumber germplasm: isozyme genetic stocks W6743, W6744, W6745

Additional index words. Cucumis sativus, allozyme, genetic markers, seed purity assessment, plant variety protection

Jack E. Staub¹, Vladimir Meglic² and Linda K. Crubaugh³

Vegetable Crops Research, U. S. Department of Agriculture, Agricultural Research Service, Department of Horticulture, University of Wisconsin, Madison, Wisconsin 53706

Received for publication______. Mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the USDA and does not imply its approval to the exclusion of other products that may be suitable.

¹ Research Horticulturist and Professor.
² Former Graduate Student.
³ Research Technician.

A series of cucumber (Cucumis sativus L. var. sativus) populations (W6743, W6744, W6745) containing alternate alleles (allozymes) for 20 enzyme coding loci were released in September 1995 by the Agricultural Research Service, United States Department of Agriculture. Lines within each population have been created through backcrossing and self-pollination to incorporate allozymes in three genetic backgrounds; European glasshouse, U.S. processing (pickling) and U.S. market (slicing) types. Within each population inbreds developed from single plant selections should be heterogeneous at specific isozyme or morphological loci, and homozygous at others. These genetic stocks could be used for genetic marker research (e.g., linkage assessment, genetic drift detection) or by breeding programs (the incorporation of unique allelic constructs into elite lines) as a tool for varietal purity assessment and plant variety protection.

Origin

Allelic variation was initially observed in a survey of elite, publicly-released, processing cucumber inbred and plant introduction accessions present in the U.S. cucumber collection (USDA Regional Plant Introduction Station, Ames IA) in adenylate kinase (AK), fructose diphosphatase (FDP), glucosephosphate isomerase (GPI), glutathione reductase (GR), glycerate dehydrogenase (G2DH), isocitrate dehydrogenase (IDH), malate dehydrogenase (MDH), manosephosphate isomerase (MPI), peptidase with glycyl-leucine (PEP-GL), peptidase with leucyl-alanine (PEP-LA), peptidase with phenylalanyl-proline (PEP-PAP), peroxidase (PER), phosphoglucomutase (PGM), 6-phosphogluconate dehydrogenase (PGD), and shikimate dehydrogenase (SKDH) (Knerr et al., 1989, Knerr et al., 1994). Some allozymes of enzymes (e.g. G2DH, IDH, MPI, PGD, PEP-LA, SKDH) were not present in elite lines and were recovered from exotic germplasm (e.g., C. sativus var. hardwickii (R) Alef.; PI 183967 and PI 215589; Table 1). The inheritance of allozymes for these enzymes conforms to Mendelian expectations, and the linkage relationships among these allozymes and between allozymes and some economically important morphological loci have been characterized (Table 2; Knerr et al., 1989; Meglic, 1994).

Crosses were made among elite lines, and between elite lines and exotic germplasm to incorporate rare allozymes into adapted germplasm. Lines (>F₄) were developed and used to determine the inheritance of the isozyme banding patterns resolvable in 15 enzyme systems using horizontal starch gel electrophoresis. These lines (5 to 10) were crossed to a European line (F₃; European glasshouse type) derived from the intermating of three proprietary glasshouse lines from Numhems Zaden BV, De Ruiter Zonen BV, and Nickerson Zwaan, BV, Poinsett 76 (U.S. market type) and GY14 (U.S. processing type). These three lines were used as recurrent parents during backcrossing in which cross progeny heterozygous for isozyme loci were identified and used as parents. After four backcrosses, lines were self-pollinated for two generations and selected for alternate allozymes at each of 20 loci (Ak-2, Ak-3, Fdp-1, Fdp-2, Gpi-1, Gr-1, G2dh, Idh, Mdh- 1, Mdh-2, Mdh-3, Mpi-1, Mpi-2, Pep-gl, Pep-la, Pep- pap, Per, Pgm, Pgd-1 and Skdh), and uniformity in the three genetic backgrounds. This resulted in the production of 6 European glasshouse (W6743), 11 U.S. processing (W6744) and 8 U.S. market type (W6745) lines which were homozygous for alternate alleles [e.g. Ak- 2 (11) and (22)] at specific loci (Table 3).

Description

Although these BC₄S₂ lines are distinct and can be placed into broad classifications according to potential horticultural utility, they are not phenotypically uniform within a specific type classification (Table 3). Lines within a specific type vary in skin texture (smooth or warty) and mature fruit color (green to orange) and length:diameter ratio (L/D; measured in mm), depending on genetic background. Lines derived using European germplasm as recurrent parents tend to be smooth-skinned, relatively long and fine-spined. Lines derived from backcrossing to U.S. processing and market lines vary in spine color (black or white) and some segregate for skin texture attributes (i.e., warts and spine thickness) and exhibit a range in L/D (2.0 to 4.8). This lack of uniformity could be due to pleiotropic effects and/or linkages between allozymes and morphological traits (Meglic, 1994).

Availability

Limited seed of individual lots of these genetic stocks and composite lots of 6743-5 are available upon written request to J. E. Staub, USDA, ARS, Department of Horticulture, University of Wisconsin, Madison, WI 53706.

Literature cited

Clayton, J. W. and D. N. Tretiak. 1972. Amine-citrate buffers for pH control in starch gel electrophoresis. J. Fish. Res. Board Can. 29:11-69-1172.
Knerr, L.D., J.E. Staub, Holder D.J., and B.P. May. 1989. Genetic diversity in Cucumis sativus L. assessed by variation at 18 allozyme coding loci. Theor. Appl. Genet. 78: 119-128.
Knerr, L.D.and J.E. Staub. 1992. Inheritance and linkage relationships of isozyme loci in cucumber (Cucumis sativus L.). Theor. Appl. Genet. 84:217-224.
Knerr, L.D., V. Meglic, and J. E. Staub. 1994. A fourth malate dehydrogenase (MDH) locus in cucumber. HortScience 30:118-119.
Market, C. L. and L. Faulhaber. 1965. Lactate dehydrogenase isozyme patterns of fish. J. Exp. Zool. 159:319-332.
Meglic, V. 1994. Inheritance and linkage relationship between biochemical and morphological loci in cucumber (Cucumis sativus L.). PhD thesis, University of Wisconsin- Madison, USA.
Richmond, R.C. 1972. Enzyme variability in the Drosophila williston group. 3. Amounts of variability in the superspecies D. paulistorum. Genetics 70:87-112.
Ridgway, G. L., S. W. Sherburne, and R. D. Lewis. 1970. Polymorphism in the esterase of Atlantic herring. Trans. Amer. Fish. Soc. 99:147-151.
Selander, R. K., M. H. Smith, S. Y. Yang, W. E. Johnson, and J. B. Gentry. 1971. Biochemical polymorphism and systematics in the genus Peromyseus. I. Variation in the old-field mouse (Peromyseus polionotus). In: Studies in genetics, Univ. of Texas Publication, Austin.

Table 1

Sources of the less common allozymes in 15 cucumber (Cucumis sativus L.) enzymes^z.

Locus^y	PI	Source
Ak-2(2)	279469	Japan
Ak-2(2)	279463	Japan
Ak-3(2)	169334	Turkey
Ak-3(2)	255937	Netherlands
Fdp-1(2)	169383	Turkey
Fdp-1(2)	192940	Peoples Republic of China
Fdp-2(1)	137851	Iran
Fdp-2(1)	164952	Turkey
Gpi-1(1)	200815	Burma
Gpi-1(1)	422192	Hungary
Gr-1(1)	109275	Turkey
G2dh(1)	285606	Poland
Idh(1)	215589	India
Idh(1)	183967	India
Mdh-1 (2)	171613	Turkey
Mdh-1 (2)	209064	United States
Mdh-2 (1)	174164	Turkey
	357835	Yugoslavia
	419214	Hong Kong
Mdh-3 (2)	255236	Netherlands
	267942	Japan
	432854	People's Republic of China
Mpi-1(2)x	176954	Turkey
Mpi-1(2)x	249562	Thailand
Pep-gl(1)	113334	Peoples Republic of China
Pep-gl(1)	212896	India
Pep-gl(2)	137851	Iran
Pep-gl(2)	212896	India
Pep-la(2)	169380	Turkey
Pep-la(2)	354952	Denmark
Pep-pap(1)	163213	India
Pep-pap(1)	188749	Egypt
Per(2)	215589	India
Pgm(2)	171613	Turkey
Pgm(2)	177364	Iraq
Pgd-1(1)	169380	Turkey
Pgd-1(1)	222782	Turkey
Pgd-1(2)	188749	Egypt
Pgd-1(2)	289698	Australia
Skdh(1)^x	302443	Taiwan
Skdh(1)^x	390952	Russia

^z Sources are only given for the less common allele of a locus used based on previous studies by Knerr et al. (1989) and Knerr and Staub (1992). There is also a common allele in addition to each of those listed above.
^y Enzyme loci designation where multiple loci of an enzyme is distinguished by hyphenated numerals and alleles are enclosed in parentheses ( ).
^x Heterogeneous population containing alternate homozygous and heterozygous genotypes.

Table 2

Enzymes assayed using specific buffer systems which provided adequate resolution of isozyme loci observed in cucumber (Cucumissativus L.).

Enzyme	Abbreviation	E.C.^z designation	Buffer^y system	No. of^x loci
Adenylate kinase	AK	2.7.4.3	S-4	2
Fructose diphosphatase	FDP	3.1.3.11	A	2
Glucosephosphate isomerase	GPI	5.3.1.9	R	1
Glutathione reductase	GR	1.6.4.2	S-4	1
Glycerate dehydrogenase	G2DH	1.1.1.29	R	1
Isocitrate dehydrogenase	IDH	1.1.1.42	S-4	1
Malate dehydrogenase	MDH	1.1.1.37	S-4	3
Manosephosphate isomerase	MPI	5.3.1.8	S-4	2
Peptidase with glycyl-leucine	PEP-GL	3.4.13.11	A	2
Peptidase with leucyl-alanine	PEP-LA	3.4.13.11	M	1
Peptidase with phenylalanyl-proline	PEP-PAP	3.4.13.11	S-4	1
Peroxidase	PER	1.11.1.7	A	1
Phosphoglucomutase	PGM	5.4.2.2	R	1
6-phosphogluconate dehydrogenase	PGD	1.1.1.43	S-4	2
Shikimate dehydrogenase	SKDH	1.1.1.25	S-4	1

^z Enzyme commission number.
^y Buffers of Clayton and Tretiak (1972), Ridgway et al. (1970), and Selander et al. (1971), Markert and Faulhaber (1965) designated as C, R, and S or M, respectively.
^x Loci designated by previous examination (Knerr and Staub, 1992) or during this survey using standard criteria and nomenclature (Richmond, 1972).

Table 3

Allozyme and morphological variation in genetic stocks of cucumber (Cucumissativus L.).

	Allelic constitution of enzyme coding loci^z																				Fruit Characteristics ^y
Identity	Ak-2	Ak-3	Fdp -1	Fdp -2	Gpi -1	Gr-1	G2dh	Idh	Mdh -1	Mdh -2	Mdh -3	Mpi -1	Mpi -2	P-gl	P-la	P- pap	Per	Pgm	Pgd -1	Skdh	Skin texture	Skin color	Spine color	L:D ratio^x
European greenhouse type
6743A	12	22	22	22	22	11	22	12	22	22	11	12	11	22	22	22	11	12	22	22	Seg	G-Y to Y-G	W	3.8-6.0
6743B	12	22	22	22	22	11	22	12	12	22	11	12	11	12	22	22	11	12	22	22	Seg	G-Y to Y-G	W	3.8-6.0
6743D	12	12	12	11	22	11	22	22	11	22	11	12	12	11	12	22	11	11	12	22	Sm	G-Y to Y	W	4.0-5.5
6743E	22	22	22	22	22	22	11	22	11	22	11	22	11	22	22	22	11	11	22	22	Sm	G-Y to Y	W	3.8-5.6
6743I	22	22	22	22	22	22	22	22	11	22	11	11	11	12	22	22	11	11	22	11	Sm	G to Y	W	5.8-6.0
6743J	22	22	22	22	22	11	22	22	11	11	11	22	11	11	22	22	11	22	22	22	Seg	Y	W	3.5-3.8
U.S. processing type
6744C	12	12	22	11	22	11	22	12	11	22	11	22	22	12	22	22	22	12	22	22	W	G to O	W	2.8-3.0
6744F	22	12	12	11	22	11	22	22	22	22	11	22	22	22	22	22	11	11	22	22	W	G-Y to Y	W	3.2-4.0
6744G	22	12	11	22	11	11	22	22	12	22	12	22	22	22	12	22	11	11	22	12	W	Y to Y-G	W	3.0-4.5
6744H	22	12	12	11	22	11	22	22	12	22	22	22	22	22	22	22	11	11	22	11	W	Y to Y-G	W	2.8-3.5
6744I	11	22	22	22	12	11	22	22	11	22	22	22	22	12	22	11	11	11	22	22	W	Y to Y-G	W	2.5-3.3
6744K	11	22	22	11	11	11	22	22	11	22	12	22	22	22	22	11	11	11	22	22	Seg	Y	W	2.4-2.8
6744L	11	22	22	11	11	11	22	22	11	22	22	22	22	11	22	11	11	11	22	22	Seg	Y-G to Y	W	2.3-2.6
6744M	11	22	22	11	22	11	22	22	11	11	11	22	11	11	22	11	11	11	22	11	Seg	G-Y to Y	W	2.2-3.2
6744P	12	11	11	11	22	11	22	22	11	22	11	11	22	11	22	11	11	11	11	22	W	Y-G to Y	W	2.7-3.5
6744S	12	12	12	11	22	11	22	22	11	22	11	12	11	11	22	11	11	11	12	22	Seg	O	B	2.6-3.8
6744T	11	12	12	11	22	11	22	22	11	22	11	12	11	11	22	11	11	11	12	22	Seg	O to YG	B	2.0-3.5
U.S. market type
6745A	22	22	11	22	22	11	12	22	12	22	12	11	12	11	22	22	11	12	22	22	Seg	O-Y to Y	Seg	3.5-4.3
6745D	22	22	12	22	22	12	22	22	11	22	11	11	11	12	22	22	11	11	22	12	Seg	O-Y to Y	Seg	3.8-4.3
6745E	12	22	12	22	22	11	22	22	11	11	22	12	11	11	22	22	11	12	12	22	Seg	G-Y to Y	W	3.3-3.8
6745F	11	22	11	22	22	11	22	22	11	22	12	11	11	11	22	22	11	12	11	22	Seg	G to Y	W	3.0-3.8
6745G	11	22	11	22	22	11	22	22	11	22	22	11	11	11	22	22	11	11	11	22	Seg	Y-G to Y	W	3.8-4.5
6745H	12	22	12	22	22	11	22	22	11	12	12	12	11	11	22	22	11	12	12	22	W	Y-G to Y	W	3.8-4.8
6745J	12	12	22	11	22	11	22	12	11	22	22	22	11	22	22	22	22	12	22	12	Seg	Y	B	3.0-3.8
6745K	11	12	11	11	22	11	22	11	11	22	12	11	22	11	22	22	11	22	22	11	Seg	O to Y	B	3.0-4.3

^z Allozymes that occur in highest frequency are given the mobility designation 100. All other alleles produce protein products with relative mobilities to allozyme 100 (mm) as follows:
Ak-2(1)-98, Ak-3(1)-98, Fdp-1(1)-96, Gpi-1(1)-98, Gr-1(1)-97, G2dh(1)-94, Idh(1)-94, Mdh-1(2)-101.5, Mdh-2(1)-98, Mdh-4(2)-102, Mpi-1(1)-96, Mp-2(2)-103, Pep-gl(1)-98, Pep-la(1)-98, Pep-pap(1)-95, Per(2)-105, Pgm(2)-102.5, Pgd-1(1)-98, Skdh(1)-98.
^ySkin texture: Sm = smooth, W = warty, Seg = segregating; Skin color: G = green, Y = yellow, O = orange; Spine color: W = white, B = black, Seg = segregating;
^xL:D = length:diameter ratio.

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