CN1190124C - Cucumis allostriploid cucumber and used for method of cucumber breeding - Google Patents

Abstract

The present invention relates to cucumis allotriploid cucumber and a method for using the cucumber in the breeding of cucumber, which is exclusively used for improving the new species of the cucumber. The cucumis allotriploid cucumber which does not have a seed is cultivated by the interspecific hybridization a novel cucumis amphidiploid species from interspecific hybridization and a cultivated common cucumber; by the back cross of the allotriploid cucumber and the cucumber, alien translocation lines with weak light resistance and root-knot nematode resistance are obtained; by the hybridization of the good alien translocation lines and female-line cucumber, a novel cucumber species with weak light resistance and root-knot nematode resistance is cultivated.

Description

The breeding method of Cucumis allotriploid cucumber

One, technical field

The breeding method of Cucumis allotriploid cucumber of the present invention belongs to the biotechnology breeding field, is exclusively used in the chromosome engineering breeding of cucumber.

Two, technical background

Patent " the Cucumis wild species are used for the method for breed cucumber " (Chinese application number 00 1 12271.1 in the application of this seminar, publication number: CN 1279009A, open day: January 10 calendar year 2001), we save by rataria, have overcome the interspecific cross obstacle between Cucumis wild species and the cultivated species first; Utilize somatic variation successfully chromosome doubling to be carried out in double haploid species hybrid, through the fertility restorer screening, cultivate first Cucumis interspecific cross amphidiploid novel species C.hytivus in the world subsequently.This amphidiploid novel species carries a complete set of chromosome set of wild species C.hystrix, but as allotetraploid, the fertility of amphidiploid novel species is lower, is unfavorable for transduceing by backcrossing utilizing the useful foreign gene in the novel species to carry out the breed improvement of cucumber.

The allotriploid cucumber is an important intermediate materials that utilizes amphidiploid novel species C.hytivus improvement cucumber variety.Because the ploidy of allotriploid cucumber has reduced, its crossability with the dliploid cucumber has obtained enhancing (Chen et al., 2001), helps by backcrossing to the common cultivation cucumber useful external source proterties that novel species carries of transduceing.

Utilize the allotriploid cucumber can also obtain some important chromosome engineering materials, as alien translocation line etc.These materials are close with cucumber chromosome (number), and the efficient that proterties shifts can be further enhanced.The breeding practice of other crop shows: alien translocation line is the desirable chromosome material (Ceoloni et al., 1996) of improvement of crop cultivar.Therefore, allotriploid cucumber and good alien translocation line subsequently also will play an important role in the breed improvement of Cucumis crop.Yet up to now, people also do not cultivate the allotriploid cucumber of Cucumis.

Three, summary of the invention

That technical problem the objective of the invention is is narrow at the hereditary basis of present cucumber resource, a little less than the resistance, and utilize the present situation of the useful proterties difficulty of external source, the breeding method of one cover Cucumis allotriploid cucumber is provided, synthetic Cucumis allotriploid does not have the seed new breeds of cucumbers, enlarge the cucumber gene pond, more extensively introduce the wild proterties of external source (gene) effectively, obtain the new breeds of cucumbers of low light tolerance, anti-root-knot nematode.

The breeding method of technical scheme Cucumis allotriploid provided by the present invention cucumber, its

Embodiment is as follows:

1, the breeding method of Cucumis allotriploid cucumber comprises:

Cucumis interspecific cross amphidiploid new species and common cultivation cucumber are carried out reciprocal cross, after the hybridization hybrid rataria is placed MS+0.2mg L ^-1Cultivate on the medium of 6-BA, obtain regeneration plant; , regeneration plant is carried out the chromosome number of meristematic cell and identify as chromosomal film-making material with tendril, obtain Cucumis allotriploid cucumber (2n=3x=26).

By the in-vitro propagate of terminal bud and axillalry bud etc., preserve and enlarge the germ plasm resource of allotriploid cucumber.The rapid propagation in vitro medium comprises, the indefinite bud medium: MS+10 μ M BA or MS+13.3 μ M BA+1.1 μ M NAA; Root media: 1/2MS+0.2mgL ^-1IBA, 0.8g L ^-1. agar, 30g L ^-1Sucrose, pH=5.8.

2, above-mentioned Cucumis allotriploid cucumber is used for the method for breed cucumber, comprising:

1) utilize allotriploid cultivating cucumber cucumber alien translocation line:

It is maternal to select above-mentioned Cucumis allotriploid cucumber to do, and female series cucumber GY-14 makes male parent, and the rataria of hybridizing is placed MS+0.2mg L ^-1Cultivate on the medium of 6-BA, obtain regeneration plant; C-is divided technology such as band, isodynamic enzyme and RAPD molecular markers for identification combine, select low light tolerance, anti-root-knot nematode proterties, obtain long fruit type alien translocation line HHZ _1-9-12(2n=14) with middle fruit type alien translocation line HHZ _1-9-8(2n=14).

2) utilize the alien translocation line of above-mentioned acquisition to cultivate new breeds of cucumbers:

Utilize the long fruit type alien translocation line HHZ of above-mentioned acquisition _1-9-12(2n=14) with middle fruit type alien translocation line HHZ _1-9-8(2n=14) with cucumber female series GY-14 carry out 2～4 generation recurrent backcrossing and selection, cultivate good new breeds of cucumbers.

The breeding method of beneficial effect Cucumis allotriploid of the present invention cucumber compared with prior art has following advantage and good effect:

The present invention utilizes unique in the world Cucumis interspecific cross amphidiploid novel species and the hybridization between the common cultivation cucumber.Beginning thus, through links such as embryo rescue, chromosome evaluation, biochemistry, molecular engineering mark and comprehensive proterties selections, obtain Cucumis allotriploid cucumber first, is a kind of intermediate materials of important chromosome engineering breeding.The alien translocation line of the Cucumis of research initiative subsequently then is the material that can directly serve the cucumber variety improvement.The present invention has important reference meanings undoubtedly to breeding practice and the theoretical research of Cucumis and even whole vegetable crop.

Owing to widened hereditary basis, Cucumis allotriploid cucumber that is obtained and new breeds of cucumbers still are that the greenhouse has all shown the not available resistivity to root-knot nematode of common cucumber at open country.Test shows that after the inoculation, the average root knot index of allotriploid cucumber is 3.8, and in the resistance, new breeds of cucumbers is on average about 4.1, in the resistance; And contrast common cucumber root knot index is 5.0, susceptible.Simultaneously, allotriploid cucumber and new breeds of cucumbers also show the better economic proterties.But about 30 of the average results of the every strain of allotriploid cucumber, heavily about 100～150 grams of fruit are about 13 centimetres, about 3 centimetres of diameter, fruit type index about 4.3; New breeds of cucumbers number of results and common cucumber are as good as, and long and short two kind fruit type is arranged, and fruit type index is 5～6.2 fruit knots are on same joint sometimes for the allotriploid cucumber, and the fruit type is neat.Fruit green, tender fruity have the lemon local flavor like cucumber when ripe approximately, do not contain any bitter taste.Preliminary attributional analysis result shows that allotriploid cucumber fruits Protein content is 0.78%, and mineral matter is 0.35%, is higher than 0.59% and 0.28% of sub-cucumber two morning respectively.All the other proterties see Table 1.

Four, description of drawings

The cultivation of Fig. 1, allotriploid cucumber and utilize it to carry out the family tree of breed cucumber

The plant of Fig. 2, amphidiploid novel species

The chromosome 2n=4x=38 of Fig. 3, amphidiploid

Fruit of Fig. 4, allotriploid cucumber (left side) and amphidiploid new species fruit (right side)

The chromosome 2n=3x=26 of Fig. 5, allotriploid cucumber

Five, embodiment

Embodiment 1 is with Cucumis interspecific cross amphidiploid novel species (Fig. 2,3) and common cucumber " two is early sub " (southern type cucumber) vernalization (vernalization in 30 days in advance of novel species seed), or take that novel species is carried out short-day and handle (illumination 8hr/ 16hr at dark night), itself and common cucumber are bloomed synchronously.Cucumber pollen is awarded on the style of interspecific cross novel species, and the embryo who gets suitable size after 20 days cultivates.Place MS to add 0.2mg L the embryo ^-1On the medium of 6-BA, successive transfer culture after 40 days, subsequently with regeneration plant intermediate house or land for growing field crops, can gather in the crops after 60 days the likeness in form corynebacterium, be about 13 centimetres, the about 3 centimetres stenospermocarpy (Fig. 4) of diameter.The chromosome of such plant is 2n=26 (Fig. 5), is the allotriploid cucumber.Resistance the analysis showed that: its low light tolerance, anti-root-knot nematode.Subsequently, by rapid propagation in vitro and preservation, obtain the allotriploid cucumber that population size enlarges, the medium of rapid propagation in vitro comprises, the indefinite bud medium: MS+10 μ M BA or MS+13.3 μ M BA+1.1 μ M NAA; Root media: 1/2MS+0.2mgL ^-1IBA, 0.8gL ^-1, agar, 30gL ^-1Sucrose, pH=5.8.

Allotriploid cucumber and cucumber female series GY-14 are hybridized, and place MS to add 0.2mgL rataria ^-1Cultivate on the medium of 6-BA and obtain regeneration plant; Get the tender tendril (being about the 1-3 millimeter) of children, carry out chromosome counting and C-and divide the analysis of band banding pattern, and analyze, filter out alien translocation line in conjunction with isodynamic enzyme and RAPD; Identify and the economic characters evaluation through resistance, filter out the alien translocation line HHZ of the good long fruit type (fruit type index 6.0) of anti-root-knot nematode (root knot index 4.1), low light tolerance (energy normal growth under the light intensity of 8000Lx) _1-9-12

Subsequently, utilize female series GY-14 and good alien translocation line HHZ _1-9-12Carry out the recurrent backcrossing in 2～4 generations, filter out long fruit type (fruit type index 6.0) new breeds of cucumbers of anti-root-knot nematode (root knot index 4.1), low light tolerance (energy normal growth under the light intensity of 8000Lx).

Embodiment 2 utilizes above-mentioned allotriploid cucumber of cultivating and cucumber female series GY-14 to hybridize, and MS is added 0.2mgL ^-1Cultivate on the medium of 6-BA and obtain regeneration plant; To utilize tendril counting chromosome, C-to divide the analysis of band banding pattern, isodynamic enzyme and RAPD to analyze and combine, filter out alien translocation line; Identify and the economic characters evaluation through resistance subsequently, filter out the good alien translocation line HHZ of the middle fruit type (fruit type index 5.0) of anti-root-knot nematode (root knot index 4.1), low light tolerance (energy normal growth under the light intensity of 8000Lx) _1-9-8

Utilize female series GY-14 and alien translocation line HHZ _1-9-8Carry out the samsara breeding in 2～4 generations, filter out the new breeds of cucumbers of the middle fruit type (fruit type index 5.0) of anti-root-knot nematode (root knot index 4.1), low light tolerance (energy normal growth under the light intensity of 8000Lx).

Table 1 application relates to the characteristic of main material

Proterties		Genotype
							Two sub-cucumber early	The allotriploid cucumber	New species	Long fruit type improvement cucumber	Middle fruit type improvement cucumber
		Chromosome number		2n＝14	2n＝26	2n＝38						2n＝14	2n＝14
Stem	Diameter (centimetre)						0.6	0.55	0.5	0.6	0.5
		Length of internode (centimetre)	10.5	5.0	6.5	10.5						10
	Number of branch						2.6	4.2	8.2	2.6	2.5
		Leaf	Petiole length (centimetre)	12.6	8.8	8.5						12.6	12.2
Leaf area (centimetre 2)	321						250.2	180	321	298
			Male flower	The first flower joint position	3.8	4.2					5	3.8	3.8
Anthocaulus length (centimetre)	2.4	1.5					0.7	2.4	2.3
				Tube length (centimetre)	0.8	0.7				0.6	0.8	0.7
Corolla length (centimetre)	2.5	1.8					1.2	2.5	2.2
				Female flower	The first flower joint position	6.2				6.8	8.5	6.2	6.1
Ovary length (centimetre)	3.8	2.5	2.0				3.8	3.0
					Corolla length (centimetre)	2.6			2.3	1.6	2.6	2.5
Fruit	Length and diameter (centimetre)	30×5	13×3				9×3	30×5					20×4
				Seed has or not	Have	Seldom			Have	Have	Have
	Seed	Length and width (centimetre)	1×0.4				0.8×0.3	0.5×0.3				1×0.4	0.9×0.4
Resistance				Nematicide	Not	Anti-			Anti-	Anti-	Anti-
	Low light tolerance	Not	Anti-				Anti-	Anti-				Anti-

Claims (3)

1, the breeding method of Cucumis allotriploid cucumber comprises:

With Cucumis interspecific cross amphidiploid novel species Cucumis hytivus Chen ﹠amp; Kirkbride and common cultivation cucumber (C.sativus L.) carry out positive and negative interspecific cross, subsequently the hybrid rataria are placed MS+0.2mg L ^-1Carry out tissue culture on the 6-BA medium, obtain species hybrid allotriploid cucumber plant (2n=26), utilize pollen to stimulate the hybrid fruit development, form no seed type new varieties.

2, the breeding method of Cucumis allotriploid cucumber according to claim 1 is characterized in that:

Terminal bud or axillalry bud with the allotriploid cucumber plant are explant, utilize rapid propagation in vitro to enlarge the population size of allotriploid, and the medium of rapid propagation in vitro comprises: indefinite bud medium, MS+10 μ M BA or MS+13.3 μ M BA+1.1 μ M NAA; And root media, 1/2 MS+0.2mgL ^-1IBA, 0.8gL ^-1. agar, 30gL ^-1Sucrose, pH=5.8.

3, the breeding method of Cucumis allotriploid cucumber according to claim 1 and 2 is characterized in that:

1) utilize allotriploid cultivating cucumber cucumber alien translocation line:

It is maternal to select above-mentioned Cucumis allotriploid cucumber to do, and female series cucumber GY-14 makes male parent, and the rataria of hybridizing is placed MS+0.2mg L ^-1Cultivate on the medium of 6-BA, obtain regeneration plant; Divide band, isodynamic enzyme and RAPD molecular markers for identification technology to combine C-, select low light tolerance, anti-root-knot nematode proterties, obtain long fruit type alien translocation line HHZ _1-9-12(2n=14) with middle fruit type alien translocation line HHZ _1-9-8(2n=14);

2) utilize the alien translocation line of above-mentioned acquisition to cultivate new breeds of cucumbers:

Utilize the long fruit type alien translocation line HHZ of above-mentioned acquisition _1-9-12(2n=14) with middle fruit type alien translocation line HHZ _1-9-8(2n=14) with cucumber female series GY-14 carry out 2～4 generation recurrent backcrossing and selection, cultivate good new breeds of cucumbers.

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