CN1895029A - Breeding method of cabbage plasmic male sterile system materials - Google Patents

Abstract

A method for selectively culturing the cytoplasmic male sterility line of Chinese cabbage RC7 includes such steps as choosing the cytoplasmic male sterility line of cabbage-type rape (or radish) RC97-1 as female parent and the selfing line of Chinese cabbage '1138' as male parent, hybridizing, choosing excellent plant as the female parent for backcrossing, and backcrossing by continuous 7 generations while choosing the high-yield and disease-resistant plants for backcrossing.

Description

A kind of selection of cabbage plasmic male sterile system materials

Technical field

The present invention relates to crop hybrid superiority and utilize technical field, particularly a kind of selection of cabbage plasmic male sterile system materials.

Background technology

Chinese cabbage (Brassica campestris L.ssp.pekinensis (Lour) Olsson) claims Chinese cabbage, die Chinakohl, bag core white dish etc. again.Belonging to the Cruciferae rape and belong to the subspecies that can form leaf-head in the rape kind, is a biennial herb plant.Originating in China, is that China's distribution is the widest, area under cultivation is maximum, the edible maximum bulk vegetable of crowd.According to the Ministry of Agriculture's statistics, national Chinese cabbage area was 2,699 ten thousand mu in 2003, accounted for 10.02% of the national vegetables gross area, and 10,197 ten thousand tons of gross yields account for 18.87% of national vegetables gross yield.Chinese cabbage group has tangible hybrid vigour, and good hybrid combination except that the hybrid advantage can improve the output, can also not need the long period to select just can be fixed to the disease-resistant gene that waits of most dominance on the first generation of hybrid generally than conventional variety volume increase about 30%.Therefore, the application of the first generation of hybrid become that domestic and international Chinese cabbage is disease-resistant, the primary measure of high-quality, high yield, stable yields breeding.

20th century the mid-80, China Chinese cabbage produces and realizes assorted optimization substantially.At present, the heterosis utilization of Chinese cabbage mainly contains two main paties: a kind of approach is the utilization of self incompatible line.Self incompatible line is extensive use on producing at present, and the self incompatible line production first generation of hybrid exists original seed production cost height, the easy decline of parents' vitality and occurs pseudostationary easily, influences problems such as first generation of hybrid quality.Second kind of approach is the utilization of male sterile line.Utilize the male sterile line production first generation of hybrid, can overcome the shortcoming of self incompatible line, the cytoplasm male sterility line high-quality first generation of hybrid that is easy to get particularly, and have the production of hybrid seeds simple, use advantages such as convenient, with low cost, emphasis and advanced subject that the person that always is the crop breeding for many years studies.

Two during the last ten years, Chinese cabbage cytoplasm male sterility line, and research is at most Polima cabbage type rape cytoplasmic male sterilty and Ogura radish cytoplasmic male sterilty the most widely both at home and abroad.The advantage of Polima cabbage type rape sterile source is that transformation is easy, keeps spectrum and recover spectrum wide, at present by transformation to multiple crop in cruciferae, its shortcoming is that fertility is subject to environmental influence, sterile degree can not reach 100%.The advantage in Ogura radish cytoplasmic sterility source is that staminody is thorough, WUHUAFEN, and sterile plant rate and sterile degree are 100%, and shortcoming is the yellow of plant seedling leaf, growth retardation, nectary is degenerated and coordinate force is poor.

Present inventor (gardening journal, 1992,19 (4): 333～340) the allo-plasm of Chinese cabbage male sterile line CMS that utilizes cabbage type rape Polima cytoplasmic sterility source to breed _3411-7(the patent No.: 92104134.9), its sterile plant rate can reach 100%, and sterile degree is more than 95%, and the advantage crossbreed of allotting with this male sterile line has been widely used in producing, but trace-pollen appears in this male sterile line regular meeting under certain temp. and humidity change condition, has influence on the purity of crossbreed; (China's Vegetable such as Sun Rifei, 1997, (4): 32～33) from the novel radish kytoplasm Chinese cabbage male sterile material NYcms of external introduction, through too much for backcross transformation, sterile strain floral organ is normal, nectary is arranged, and it is normal to set seeds, but along with the increase of transformation algebraically, it is very fast to degenerate, part seedling performance deformity, hybrid vigour is not strong simultaneously, coordinate force is poor, can't use; (China's Vegetable, 1994, (4): 4～6) with Wang Zhao carrot male sterility and process such as deer hero ⁶⁰The low generation Chinese cabbage crossing of Co gamma-radiation has been bred 134 male sterile line of cabbage, but because of a little less than the plant strain growth gesture, coordinate force is not strong etc. former thereby fail to use; (China's Vegetable such as Wei Baoqin, 1995, (5): 18～21) utilize the Ogrua sterile source, painstaking efforts by more than 10 years, study intensively, with the Ogrua sterile source successfully transformation to Chinese cabbage, selected sterile plant rate and sterile degree and all reached 100% two Chinese cabbage male sterile line 1A1 and 1A3, but do not seen the report that crossbreed is applied to produce so far; (Shanghai Agricultural journal such as Yu Peitao, 2000,16 (1): 17～19) pass through gene engineering method, with the Agrobacterium tumefaciems plasmid is carrier, carry male sterile gene and anti-kanamycin gene as reporter gene, in addition the anti-rifampin gene that carries with this Agrobacterium is as the screening and culturing of Agrobacterium, is acceptor with the stem apex of Chinese cabbage seedling, change male sterile gene over to Chinese cabbage, but do not see and bring out the report that crossbreed is applied to produce; (North China agronomy newspaper such as Zhang Deshuan, 2002,17 (1): 60～63) to introduce the novel cabbage type rape cytoplasmic male sterilty source CMS96 of France, transformation becomes 3 parts of male sterile lines such as 01-646,01-644 and 01-709 on Chinese cabbage, though report that the sterility of these male sterile lines is stable, the hybrid combination volume increase of preparation is obvious, but does not also see the report that crossbreed is applied to produce.

Summary of the invention

Defective or deficiency at above-mentioned prior art existence, the object of the present invention is to provide a kind of selection of cabbage plasmic male sterile system materials, this method can keep the advantage of Ogura radish cytoplasmic sterility source male sterile line of cabbage sterility, can overcome defectives such as its plant leaf yellow, growth retardation, nectary degeneration and coordinate force difference again.

For achieving the above object, the technical solution adopted in the present invention is: a kind of selection of cabbage plasmic male sterile system materials, it is characterized in that, this method is a female parent with cabbage type rape radish cytoplasmic male sterilty material RC97-1, with Chinese cabbage inbred lines 1138 is male parent, obtain transformation generation seed by interspecific cross, the performance nothing similar to the transformation male parent of selecting and remain loses the backcross female parent of the fine individual plant of flower bud as next round; Through continuous 7 generation backcross transformation, and through economic characters select, disease resistance identifies, it is stable to select sterility, sterile plant rate and sterile degree reach 100%, the on all four cabbage plasmic male sterile system materials RC of economic characters and corresponding maintenance line ₇

Cabbage plasmic male sterile system materials RC of the present invention ₇, it is stable to have sterility, and sterile plant rate and sterile degree are 100%; It is healthy and vigorous to grow, and does not have and loses flower bud, and comprehensive disease resistance is strong, seedling not yellow under cryogenic conditions; Nectary development is good, and nectar is more, and column cap is normal, and blossoming and bearing fruit property is good; Coordinate force is good; Hybrid seeding, the seed production advantages of higher has higher value.

Description of drawings

Fig. 1 is cabbage plasmic male sterile system materials RC of the present invention ₇Compare with corresponding maintenance line flower, a left side is a male sterile line, and the right side is a maintenance line;

Fig. 2 is cabbage plasmic male sterile system materials RC of the present invention ₇And corresponding maintenance line spray relatively, and a left side is a male sterile line, and the right side is a maintenance line;

Fig. 3 is cabbage plasmic male sterile system materials RC of the present invention ₇The plant feature.

The specific embodiment that provides below in conjunction with the inventor comes the present invention is described in further detail.

Embodiment

1. cabbage plasmic male sterile system materials RC ₇Seed selection

RC97-1 is a female parent with cabbage type rape radish cytoplasmic male sterilty material, with Chinese cabbage inbred lines 1138 is male parent, obtain transformation generation seed by interspecific cross, the performance nothing similar to the transformation male parent of selecting and remain loses the backcross female parent of the fine individual plant of flower bud as next round; Through continuous 7 generation backcross transformation, and through economic characters select, disease resistance identifies, it is stable to select sterility, sterile plant rate and sterile degree reach 100%, the on all four cabbage plasmic male sterile system materials RC of economic characters and corresponding maintenance line ₇

Above-mentioned male parent Chinese cabbage inbred lines 1138 derives from the middle Indian mallow leaf Chinese cabbage that introduce in the Inner Mongol, separates through inbreeding of more generation, according to a conventional method the systematic breeding gained.Its plant profile is: the lotus throne leaf is upright, wealthy lanceolar, and dark green leaf color, leaf margin is wavy, the blade face shrinkage, vein is fine and closely woven, and middle rib is generous, light green, the high stake of leaf-head type, plant height 58.6cm, strain width of cloth 45.3cm, the long 56.6cm of leaf, the wide 29.7cm of leaf, 13 of the outer numbers of sheets, the vertical footpath of leaf-head 48.5cm, transverse diameter 16.3cm, spherical index 2.98, about the heavy 3.6kg of single ball, growth is neat, and comprehensive disease resistance is strong.

1.1 the backcross transformation of sterile material

Since spring in 1997, (the cabbage type rape radish cytoplasm male sterility line is French scholar Bannerot to the present inventor at the cabbage type rape radish cytoplasmic male sterilty material RC97-1 to introduction, H. equal 1974 with in the European cabbage type rape cell nucleus importing japanese radish sterile cytoplasm, through the continuous backcross transformation, bred cabbage type rape radish cytoplasmic male sterile line, this male sterile line is sterile thoroughly, sterility is insensitive to environment reaction, nearly all cabbage type rape variety can both be made its maintenance line, and transformation becomes different ripe property easily, the male sterile line of different qualities is acknowledged as the rape sterile material that sterility is the most stable in the world.Oil crop research institute of the Chinese Academy of Agricultural Sciences 1980, nineteen eighty-two are carried out transformation after introducing this sterile material from France, Canada respectively on different cabbage type rapes.Academy of agricultural sciences, Shaanxi Province economic crops research institute nineteen eighty-two is introduced the sterile material of this transformation by Chinese oil plant research institute, continuation transformation on different cabbage type rapes) floral organ structure, sterile proterties etc. are observed evaluation, confirmed that this material is sterile complete, behind the stable cytoplasmic male sterilty material of proterties, select its growth potential strong, nectary development is normal, do not have and lose flower bud, the complete sterility individual plant that the cross-fertile rate is good is maternal, priority is dissimilar with the applicant's seed selection for many years, 17 parts of Chinese cabbage parent materials that coordinate force is good are male parent, adopt artificial florescence bagging pollination technique, carry out the hybridization of male sterile line RC97-1 and dissimilar Chinese cabbage inbred lines and many generations and backcross its sterility of transformation.Method is: flowering stage, pollen with 17 parts of good Chinese cabbage strains is applied on the column cap of corresponding male sterile line RC97-1 respectively, obtain transformation generation seed, being carried out to strain phase economic characters autumn selects, the botany proterties of investigation plant, superseded petiole has the intermediate materials such as decomposite leaf type of incising, being partial to cabbage type rape, and the performance nothing similar to the transformation male parent of selecting and remain loses the backcross female parent of the fine individual plant of flower bud as next round.Simultaneously, its corresponding samsara male parent is carried out the selfing purifying.In order to accelerate transformation speed, taked greenhouse, open country and strange land to add the method that combines for transformation, specific practice is: backcross maternal and the samsara male parent was carried out open field live broadcast in about August 10 to selected, in 11 months, become the strain phase last ten-days period, the selection of economic characters is carried out in strictness, selected kind strain is colonizated in the greenhouse, the temperature in greenhouse should remain on 20 ℃～25 ℃, and give sufficient illumination, generally by the end of December to mid-January bolting bloom, can carry out artificial back pollinating with the samsara male parent, can receive seed to late Febuary.With the seed accepted 22 ℃～25 ℃ following vernalization, when treating that seed is sprouted the dew bud, put it in 2 ℃～3 ℃ refrigerators, carried out incomplete vernalization 20 days, and in late March is seeded into 22 ℃～25 ℃ greenhouse, grew seedlings, in 4 months, adopt the film-mulching cultivation mode last ten-days period, field planting is in the area that cools of height above sea level 1300m～1500m, generally late May to mid-June, bolting was bloomed, and can proceed artificial back pollinating with the samsara male parent, and late July can be received seed.Like this, can guarantee to add for twice of transformation in 1 year.When backcrossing for 7 generations, the male sterile line germplasm materials economic characters that obtain are with samsara male parent (maintenance line) consistent (Fig. 1,2).

1.2 economical character is observed

Each transformation is from generation to generation in becoming strain phase to observe the economical characters such as leaf, leaf look, sphere, plant height, the strain width of cloth, the bright sample quality of individual plant of the sterile material and the male parent of backcrossing thereof.Florescence check sterile material corolla, pattern, stamen, gynoecium and nectary size, shape and lose flower bud situation etc.After the artificial pollination, when kind of pod is ripe, observe its situation that bears pods, comprise that the sum that bears pods, pod length, pod slightly reach every pod seed number etc.And each from generation to generation from early flowering season to the whole florescence, the stamen of checking sterile material whether degenerate fully and flower pesticide in whether have pollen.Table 1 is transformation during 7 generations, and the Other Main Agronomic Characters of the corresponding transformation male parent with it of all types of Chinese cabbage materials of transformation relatively.

The Chinese cabbage male sterile material in 7 generations of table 1 transformation and the Other Main Agronomic Characters of corresponding transformation male parent thereof are relatively

Test material	Leaf	The leaf look	Spherical	Plant height/cm	The strain width of cloth/cm	Maximum lotus throne leaf		The bright sample quality/kg of individual plant
									Length/cm	Wide/cm
						RC 1	Nearly circle				Green	Spherical	35.3	35.6	29.3	25.4	2.7
B 1	Nearly circle	Green	Spherical	27.5	30.2			25.1	21.2	1.9
						RC 2	Nearly circle				Light green	Spherical	37.8	37.6	31.4	27.6	2.8
B 2	Nearly circle	Light green	Spherical	28.4	34.5			26.6	23.9	2.1
						RC 3	Wide ovum				Green	Head shoot shape	39.4	43.4	29.5	25.6	3.2
B 3	Wide ovum	Green	Head shoot shape	35.6	40.2			27.5	23.3	2.2
						RC 4	Oval				Light green	Tubular	46.7	40.4	38.8	20.1	2.5
B 4	Oval	Light green	Tubular	47.2	38.0			36.5	21.4	2.4
						RC 5	Long ovum				Dark green	Shell shape	38.9	39.3	37.6	22.3	2.8
B 5	Long ovum	Dark green	Shell shape	34.0	33.7			34.8	18.4	1.8
						RC 6	Fall ovum				Green	Head shoot shape	42.6	54.5	38.8	27.3	4.6
B 6	Fall ovum	Green	Head shoot shape	40.8	52.4			36.4	24.5	3.6
						RC 7	Oval				Green	Tubular	58.6	45.3	56.6	29.7	3.6
B 7	Oval	Green	Tubular	58.4	44.5			56.0	29.5	3.6
						RC 8	Oval				Light green	Tubular	60.9	52.9	57.3	29.8	4.4
B 8	Oval	Light green	Tubular	55.3	46.5			54.2	26.5	3.9
						RC 9	Fall ovum				Light green	Head shoot shape	43.2	44.8	39.4	29.7	3.6
B 9	Fall ovum	Light green	Head shoot shape	40.6	40.3			36.4	26.8	2.5
						RC 10	Nearly circle				Light green	Spherical	43.5	44.9	34.6	29.3	3.3
B 10	Nearly circle	Light green	Spherical	37.2	39.4			32.2	28.4	2.3
						RC 11	Oval				Dark green	Tubular	56.7	52.9	56.8	29.6	4.3
B 11	Oval	Dark green	Tubular	52.3	48.2			52.1	26.6	3.5
						RC 12	Nearly circle				Green	Spherical	44.3	50.6	35.6	33.2	3.4
B 12	Nearly circle	Green	Spherical	38.4	45.1			33.1	30.0	2.8
						RC 13	Long ovum				Green	Tubular	57.4	58.5	55.5	30.7	3.6
B 13	Long ovum	Green	Tubular	52.0	52.8			52.9	27.5	2.5
						RC 14	Long ovum				Green	Shell shape	50.5	55.9	52.6	30.5	4.1
B 14	Long ovum	Green	Shell shape	45.6	47.8			47.2	27.6	2.6
						RC 15	Long ovum				Green	Shell shape	53.8	49.5	53.4	30.3	3.9
B 15	Long ovum	Green	Shell shape	47.2	44.4			46.3	27.9	2.6

RC 16	Long ovum	Green	Head shoot shape	53.8	56.5	52.5	30.0	4.7
									B 16	Long ovum	Green	Head shoot shape	49.3	50.7	47.2	27.4	3.2
RC 17	Wide ovum	Light green	Spherical	36.7	40.6	34.2	33.2	2.8
									B 17	Wide ovum	Light green	Spherical	36.1	40.2	32.7	29.8	2.3

Annotate: " RC " represents sterile material in the table, and " B " represents corresponding transformation male parent (being maintenance line)

Select through the backcross transformation in 7 generations and strict economic characters, in success in 2003 select that sterility is stable, sterile plant rate and sterile degree reach 100%, economic characters and corresponding maintenance line are in full accord, and growth is neat, can be used for the cabbage plasmic male sterile system materials RC of formulated in combination ₇

1.3 male sterile line germplasm materials RC ₇The principal character characteristic

1.3.1 floral organ feature

Male sterile line germplasm materials RC ₇Flower compare with maintenance line, petal is slightly littler than corresponding maintenance line, 6 pieces on stamen, milky, flower pesticide is shrivelled, white, fine, be degenerated to sarconeme shape, stamen and flower pesticide length obviously shorten, and examine under a microscope, and can not find the pollen of this male sterile line; Gynoecium Dan Sheng, the many mastoid processes of stigma surface, ovary mode of appearance size is basic identical with maintenance line, and structure, function are normal; Nectary quantity is identical with maintenance line, and development condition is good, and look green, and the tool fragranced, can attract the normal gathering honey pollination of insect (table 2) such as honeybee.

Table 2 male sterile line new germ plasm material RC ₇Compare with maintenance line floral organ feature

Test material	Lose flower bud	Corolla			Stamen	Gynoecium		Nectary
									Degree of development/mm	Petal size/mm	Color	Shape	Length/mm
		RC 7	Do not have	12.2× 11.5		8.5× 5.0	Deep yellow							Milky, sarconeme shape, flower pesticide is shrivelled, fine, WUHUAFEN	Directly	6.1	Normally
B 7	Do not have				13.8× 12.7			9.4× 6.5	Deep yellow	Milky, thread, flower pesticide be normal, flower pesticide is arranged	Directly	6.3	Normally

Annotate: this table is survey data in 2005

1.3.2 male sterile line germplasm materials RC ₇Sterility performance

1.3.2.1 Temperature Influence

In the period of 2003～2006, continuous 4 years in winter, spring in greenhouse and land for growing field crops, and planted in different ecological areas is to male sterile line RC ₇Sterility observe evaluation, the result is male sterile line RC ₇Sterility stable, sterile plant rate reaches 100%, sterile degree also is 100%.

1.3.2.2 the influence of different sowing dates

In the spring 2004 Winter Solstice in 2003, divide December 20, January 5, January 20 and four date of seeding sowings on February 5 male sterile line RC ₇, be colonizated in the land for growing field crops on March 25, sterile plant rate of investigation in 4 days and individual plant sterile degree from every interval of early flowering season.The investigation result of 4 dates of seeding is: male sterile line RC ₇Sterile plant rate be 100%, sterile degree is 100% also, and male sterile line RC under the different sowing dates condition is described ₇Sterility be not subjected to the influence of environmental condition.

1.3.3 male sterile line germplasm materials RC ₇Morphological observation

Spring in 2006 is to male sterile line germplasm materials RC ₇Carried out cytomorphology research, the result shows: male sterile line germplasm materials RC ₇Anther development is hampered by tetrad to the monokaryon pollen period, and flower pesticide vacuole occurs in the early stage tapetal cell of reduction division, and central authorities form one at the flower blister cavities, cause microspore development to be obstructed, and form cavity flower pesticide.

1.3.4 comprehensive gardening proterties

Select through the backcross transformation in 7 generations and strict economic characters, on the main plant proterties, the male sterile line RC that breeds ₇Economic characters and corresponding maintenance line B ₇Basically identical (Fig. 2).85 days breeding times, plant height 58.6cm, strain width of cloth 45.3cm, the leaf look dark green, and leaf margin is wavy, the high stake of leaf-head type, the long 56.6cm of leaf, the wide 29.7cm of leaf, 13 of the outer numbers of sheets, the vertical footpath of leaf-head 48.5cm, transverse diameter 16.3cm, spherical index 2.98, about the heavy 3.6kg of single ball, growth is neat, comprehensive disease resistance strong (table 3).

Table 3 male sterile line new germ plasm material RC ₇Compare with the comprehensive gardening proterties of maintenance line

Test material	Plant height (cm)	The strain width of cloth (cm)	Leaf long (cm)	Leaf wide (cm)	The outer number of sheets (sheet)	The vertical footpath of leaf-head (cm)	Leaf-head transverse diameter (cm)	Spherical index	Single ball heavy (kg)	Breeding time (d)
											RC 7	58.6	45.3	56.6	29.7	13	48.5	16.3	2.98	3.6	85
B 7	58.4	44.5	54.0	29.6	13	48.5	16.5	2.94	3.6	85

1.3.5 solid ability

1.3.5.1 compare with corresponding maintenance line fecundity

The continuous result of the test in 2 years showed in 2004～2005 years: male sterile line RC ₇With maintenance line B ₇In 3: 1～4: 1 ratio field planting in the gauze canopy, florescence artificial supplementary pollination, male sterile line RC ₇Pod long, single pod is set seeds and maintenance line B ₇No significant difference (table 4).

Table 4 male sterile line new germ plasm material RC ₇Compare with the fecundity of maintenance line

Test material	2004			2005
							Pod long (cm)	Single pod set seeds (grain)	Thousand kernel weight (g)	Pod long (cm)	Single pod set seeds (grain)	Thousand kernel weight (g)
	RC 7	5.8	20.2	2.3	5.2	16.4							3.2
B 7							5.8	20.4	2.1	5.0	13.8	3.0

1.3.5.2 the solid situation of hybrid combination spontaneous pollination

Use male sterile line RC ₇The hybrid combination RC of preparation ₇* 04S ₂₄₅And RC ₇* 03S ₁₂₀₇2005～2006 years in natural isolated area by the honeybee natural pollination, by inquiry, the cross-fertile rate of two hybrid combinations is high, reaches 100%, wherein, single pod is set seeds to count and can reach 21 at most, minimum is 9, and seed collecting strain average branch number (1,2 grade) is respectively 35.8 and 37.3, and the individual plant seed yield is respectively 42.6g and 44.2g (Fig. 3).This shows male sterile line RC ₇Nature fecundity good (table 5).

Table 5 male sterile line new germ plasm material RC ₇The solid situation of spontaneous pollination

Test material	Plant height (cm)	Stem thick (cm)	Branch amount (bar)		Single-strain legumen number (individual)	Single pod set seeds (grain)	Thousand kernel weight (g)	Individual plant seed yield (g)
									1 grade	2 grades
			RC 7×04S 245	118.5							1.4	11.5	24.3	894.8	19.4	3.6	42.6
RC 7×03S 1207	123.7	2.1			12.8	24.5	962.5	19.2	3.8	44.2

1.3.6 disease resistance qualification result

1.3.6.1 inoculate disease resistance and identify artificial seedling stage

2004 to male sterile line RC ₇And maintenance line B ₇Carrying out inoculating artificial seedling stage disease resistance identifies.The result is: male sterile line RC ₇Virus disease disease index 10.8 shows as high anti-(HR); Downy mildew disease index 9.9 shows as high anti-(HR) (table 6).

Table 6 male sterile line new germ plasm material RC ₇Artificial disease resistance qualification result with maintenance line

Expert evidence	Repeat	Virus disease		Downy mildew
						Disease index	Disease-resistant classification	Disease index	Disease-resistant classification
		RC 7	I	7.8	HR					5.1	HR
II	14.4					R	11.4	R
			III	10.3	HR				13.2	R
On average	10.8					HR	9.9	HR
			B 7	I	9.4				HR	7.8	HR
II	13.7	R				14.8	R
				III	11.5			R	13.7	R
On average	11.5	R				12.1	R

Annotate: HR: expression is high anti-, R: represent disease-resistant

1.3.6.2 field resistance is identified

2004 to male sterile line RC ₇And maintenance line B ₇Carrying out field resistance identifies.The result is: male sterile line RC ₇Virus disease disease index 11.2 shows as disease-resistant (R); Downy mildew disease index 8.8 shows as high anti-(HR); Black spot disease index 19.2 shows as disease-resistant (R); The soft rot incidence of disease is 0 (table 7).

Table 7 male sterile line new germ plasm material RC ₇With maintenance line field resistance qualification result

Expert evidence	Repeat	Virus disease		Downy mildew		Black spot		The soft rot incidence of disease (%)
									Disease index	Disease-resistant classification	Disease index	Disease-resistant classification	Disease index	Disease-resistant classification
		RC 7	I	13.5	R	5.7	HR								18.9	R	0
II	8.2							HR	11.5	R	23.0	R	0
			III	11.8	R	9.2	HR							15.8	R	0
On average	11.2							R	8.8	HR	19.2	R	0
			B 7	I	15.7	R	6.9							R	14.6	R	0
II	12.5	R						7.2	R	22.8	R	0
				III	13.4	R	12.5						R	15.3	R	0
On average	13.9	R						8.9	HR	17.6	R	0

Annotate: HR: expression is high anti-, R: represent disease-resistant

2. Chinese cabbage cytoplasm male sterility line new germ plasm material RC ₇Breeding

Use Chinese cabbage cytoplasm male sterility line RC ₇Be female parent, with corresponding maintenance line B ₇For male parent is hybridized, breeding Chinese cabbage cytoplasm male sterility line new germ plasm material RC ₇Concrete steps are: before autumn Chinese cabbage results, in strict accordance with parent's standard male sterile line RC that selects and remain ₇With maintenance line B ₇Maternal plant is heeled in, safe overwintering, and at the beginning of 3 months spring of next year, field planting is in 40～60 order gauze isolated areas, and plantation density is line-spacing 60cm, spacing in the rows 40cm.

Male sterile line RC ₇Breeding canopy: male sterile line RC ₇With maintenance line B ₇In ratio field planting in 4: 1～5: 1, fixedly special messenger's pollination, or manually put honeybee in the net canopy, with maintenance line B ₇Pollen is authorized male sterile line RC ₇, in time pull out maintenance line B after the blossom fall ₇Plant is planted strain and gathers stage of yellow ripeness.

Maintenance line B ₇The breeding canopy: artificial florescence supplementary pollination, or manually put honeybee in the net canopy, self propagated is planted strain and is gathered stage of yellow ripeness.

Claims (4)

1. the selection of a cabbage plasmic male sterile system materials, it is characterized in that, this method is a female parent with cabbage type rape radish cytoplasmic male sterilty material RC97-1, with Chinese cabbage inbred lines 1138 is male parent, obtain transformation generation seed by interspecific cross, the performance nothing similar to the transformation male parent of selecting and remain loses the backcross female parent of the fine individual plant of flower bud as next round; Through continuous 7 generation backcross transformation, and through economic characters select, disease resistance identifies, it is stable to select sterility, sterile plant rate and sterile degree reach 100%, the on all four cabbage plasmic male sterile system materials RC of economic characters and corresponding maintenance line ₇

2. the method for claim 1 is characterized in that, the plant profile of described Chinese cabbage inbred lines 1138 is: the lotus throne leaf is upright, wealthy lanceolar, dark green leaf color, leaf margin is wavy, the blade face shrinkage, vein is fine and closely woven, middle rib is generous, light green, the high stake of leaf-head type, plant height 58.6cm, strain width of cloth 45.3cm, the long 56.6cm of leaf, the wide 29.7cm of leaf, 13 of the outer numbers of sheets, the vertical footpath of leaf-head 48.5cm, transverse diameter 16.3cm, spherical index 2.98, about the heavy 3.6kg of single ball, growth is neat, and comprehensive disease resistance is strong.

3. the method for claim 1 is characterized in that, described cabbage plasmic male sterile system materials RC ₇Feature be:

1. floral organ feature

Chinese cabbage cytoplasm male sterility line RC ₇Flower compare with maintenance line, petal is slightly littler than corresponding maintenance line, 6 pieces on stamen, milky, flower pesticide is shrivelled, white, fine, be degenerated to sarconeme shape, stamen and flower pesticide length obviously shorten, and examine under a microscope, and can not find the pollen of this male sterile line; Gynoecium Dan Sheng, the many mastoid processes of stigma surface, ovary mode of appearance size is basic identical with maintenance line, and structure, function are normal; Nectary quantity is identical with maintenance line, and development condition is good, and look green, and the tool fragranced, can attract the normal gathering honey pollination of insect such as honeybee;

2. comprehensive gardening proterties

85 days breeding times, plant height 58.6cm, strain width of cloth 45.3cm, the leaf look dark green, and leaf margin is wavy, the high stake of leaf-head type, the long 56.6cm of leaf, the wide 29.7cm of leaf, 13 of the outer numbers of sheets, the vertical footpath of leaf-head 48.5cm, transverse diameter 16.3cm, spherical index 2.98, about the heavy 3.6kg of single ball, growth is neat, and comprehensive disease resistance is strong

4. the method for claim 1 is characterized in that, described cabbage plasmic male sterile system materials RC ₇Propagation method, adopt Chinese cabbage cytoplasm male sterility line RC ₇Be female parent, with corresponding maintenance line B ₇Hybridize for male parent, concrete steps are: before autumn Chinese cabbage results, in strict accordance with parent's standard male sterile line RC that selects and remain ₇With maintenance line B ₇Maternal plant is heeled in, safe overwintering, in field planting at the beginning of 3 months spring of next year in 40～60 order gauze isolated areas, male sterile line RC ₇With maintenance line B ₇In ratio field planting in 4: 1～5: 1, plantation density was line-spacing 60cm, spacing in the rows 40cm, and fixedly special messenger's pollination of florescence, or manually put honeybee in the net canopy, with maintenance line B ₇Pollen is authorized male sterile line RC ₇, in time pull out maintenance line B after the blossom fall ₇Plant is planted strain and gathers stage of yellow ripeness.

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