CN107278879B - Method for cultivating nutrient-rich loose cauliflower - Google Patents
Method for cultivating nutrient-rich loose cauliflower Download PDFInfo
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- A—HUMAN NECESSITIES
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- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
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Abstract
The invention discloses a method for cultivating nutrient-rich loose cauliflower. The invention discloses a method for cultivating nutrient-rich loose cauliflower, which comprises the following steps: hybridizing white cauliflowers respectively named as white cauliflower 1 and white cauliflower 2 to obtain a first filial generation; white cauliflower 1 is white loose cauliflower, and white cauliflower 2 is white cauliflower cytoplasmic male sterile material; backcrossing the first filial generation serving as a female parent and the white cauliflower 1 serving as a recurrent parent with the first filial generation, and screening to obtain backcross progeny of cytoplasmic male sterility; and (4) hybridizing the backcross progeny serving as a female parent and the purple cauliflower serving as a male parent, and screening to obtain the nutrient-rich loose cauliflower. Experiments prove that the nutritional quality and the edible quality of the nutrient-rich loose cauliflower obtained by the method are obviously improved, and the market prospect is wide.
Description
Technical Field
The invention relates to a method for cultivating nutrient-rich loose cauliflower, belonging to the field of plant breeding.
Background
Cauliflower, also known as cauliflower, broccoli flower and the like, is a variety which can form a cauliflower bulb in brassica species in cruciferae, and is planted in large area and widely consumed in various places in China. The edible organs are flower balls, which are mostly milk white, and also have various variation types such as purple, orange, light green and the like. Due to the taste and consumption habits, the cauliflowers produced and consumed in China are almost white ball-shaped and basically compact ball-shaped.
With the improvement of living standard, people are more and more pursuing the flavor, taste and nutritional quality of vegetables. The consumption demand of cauliflower is also changing, and loose cauliflower with better mouthfeel and quality is more popular in recent years. The loose cauliflower has long pedicel, crisp and tender mouthfeel and good quality, is very popular with consumers, has higher planting yield than the traditional compact cauliflower, is very popular with growers, and is rapidly developed to a large-area loose cauliflower production base in south and north China from Fujian and Zhejiang in short years. In recent years, purple cauliflower is also planted in a small amount, but the planting scale is limited due to the fact that the purple cauliflower tastes hard and tight.
Disclosure of Invention
The invention aims to cultivate the loose cauliflower rich in nutrition.
The invention firstly provides a method for cultivating nutrient-rich loose cauliflower, which comprises the following steps:
A1) hybridizing white cauliflowers respectively named as white cauliflower 1 and white cauliflower 2 to obtain a first filial generation;
the white cauliflower 2 is a white cauliflower cytoplasmic male sterile material;
A2) taking the first filial generation of A1) as a female parent, taking the white cauliflower 1 as a recurrent parent, carrying out backcross with the first filial generation, and screening to obtain a backcross progeny of cytoplasmic male sterility;
A3) hybridizing the backcross progeny serving as a female parent and the purple cauliflower serving as a male parent, and screening to obtain the nutrient-rich loose cauliflower;
the nutrient-rich bulk cauliflower has increased nutritional value compared to the white cauliflower 1.
The white cauliflower 1 was used as a male parent in a1) and the white cauliflower 2 was used as a female parent in a 1).
In the above method, the increase in the nutritional value may be expressed in 1) and/or 2) below:
1) the nutrient-rich bulk cauliflower anthocyanidin, vitamin C, titratable acid, soluble solids, crude fiber, protein, mineral and/or glucosinolate components beneficial to the human body are increased compared to the white cauliflower 1;
2) compared with the white cauliflower 1, the nutrient-rich loose cauliflower has reduced content of glucosinolate which is not beneficial to human body.
In the above method, the mineral may be calcium, potassium, magnesium, sodium, phosphorus, copper, iron, manganese and/or zinc.
The thioglucoside component beneficial to human body can be RAA, NAP, 4OH, GBC, 4ME and/or NEO.
The glucosinolate component not beneficial to human body can be PRO.
In the above method, the white cauliflower 1 may be a loose type cauliflower, and the white cauliflower 2 provides a cytoplasmic male sterile source, and may be a loose type or a compact type cauliflower.
The purple cauliflower can be purple compact cauliflower.
In the above method, the backcross in A2) may be a series of 5-7 backcrosses. A2) The backcross in (1) may specifically be a continuous 5-generation backcross.
The backcross progeny may be loose cauliflower.
In the above method, the white cauliflower 1 may have the following properties: strong growth potential, moderate plant type, good disease resistance, loose flower ball, white color and/or long flower stalk.
In the above method, the white cauliflower 2 may have the following properties: cytoplasmic male sterility, strong growth potential, moderate plant type, good disease resistance and/or white color.
In the above method, the purple cauliflower may have the following properties: strong growth potential, moderate plant type, good disease resistance and/or purple flower ball.
In the above method, the white cauliflower 1 may be a loose cauliflower variety Qingnong 90 or a homozygous progeny thereof.
The white cauliflower 2 may be Jingyan 50 or its progeny. The Jingyan 50 may be Jingyan Yinong (Beijing) science and technology company Limited.
The purple cauliflower can be purple crystal or homozygous progeny thereof. The amethyst can be amethyst of the research institute of new country and wild variety in Henan.
In the above method, the white cauliflower 1 may be a homozygous plant obtained by culturing free microspores of a loose cauliflower variety Qingnong 90.
In the above method, the purple cauliflower may be a homozygous plant obtained by culturing purple crystals with free microspores.
The method for cultivating the nutrient-rich loose cauliflower creates the high-nutrient purple loose cauliflower rich in glucosinolate, anthocyanin, Vc and mineral substances, and adds a high-quality cauliflower variety with high nutrient and health-care values to the 'vegetable basket' in China. The nutrient-rich loose cauliflower provided by the invention has the following outstanding advantages:
(1) the obtained nutrient-rich loose cauliflower has the basic characteristics of common white loose cauliflower, namely long and crisp pedicel and loose curd, commercial organs (curd, namely edible organs) are purple, and the nutrient-rich loose cauliflower is rich in main nutrient health-care components such as Vc, protein, mineral nutrition, anthocyanin, glucosinolate (RAA, NAP, 4OH, GBC, 4ME and NEO) and reduces the content of glucosinolate PRO which is not beneficial to human health, wherein the PRO is proved to be a precursor of goitrogen 5-ethylene oxazolidine-2-thione, and the content is high and is not beneficial to human health;
(2) compared with the common white loose cauliflower, the loose cauliflower has the advantages that the nutritional quality is remarkably improved, the main nutritional ingredients are remarkably improved, and particularly, the anthocyanin which can remove free radicals of a human body and has a strong anti-oxidation function is added;
(3) compared with the common purple cauliflower, the loose cauliflower has longer flower stalks, is crisp and tender, has loose flower balls, obviously improves the eating quality, has the taste of white loose cauliflower, has crisp, tender and slightly sweet flower stalks, and is more popular with consumers;
(4) the loose cauliflower is prepared by using a cytoplasmic male sterile line, the sterile line seedlings are not yellowed at low temperature, the floral organs are normal, the honey glands are developed, the flower honey is more, the sterile plant rate and the sterile degree of the sterile plant reach 100%, the hybridization rate can reach 100%, and the seed quality is high.
The breeding method provided by the invention is simple and convenient, and the bred loose cauliflower has the advantages of excellent quality, high nutrition and health care value and wide market prospect.
Drawings
FIG. 1 shows a process for cultivating a nutrient-rich loose cauliflower.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The experimental procedures in the following examples are conventional unless otherwise specified. Materials, reagents, instruments and the like used in the following examples are commercially available unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
Example 1 cultivation of nutrient-rich Loose cauliflower
First, the breeding process
The breeding process is shown in FIG. 1.
1. White loose cauliflower Qingnong 90 (available in the market) is purchased in the market, a large number of regeneration homozygous plants are obtained by utilizing a free microspore culture technology, 10 excellent single plants with strong growth potential, moderate plant type, good disease resistance, loose and white flower balls and longer flower stalks are screened out through field comprehensive character investigation and identification, and 10 excellent loose cauliflower homozygous lines are obtained through respective single plant selfing. Through combining ability determination and nutrition quality analysis, 1 loose type cauliflower homozygous strain (S5, white cauliflower 1) with strong combining ability and high content of nutrient components is screened out.
Main characteristic features of S5: the plant is semi-upright, the plant height is 50cm, the development degree is 55 multiplied by 60cm, 15 outer leaves are adopted, the plant is oval, light green, the leaf surface wax powder is medium, the flower ball is flat, round, loose and white, the flower stalk is 14cm long, the weight of a single ball is 0.7Kg, the ball height is 13cm, the ball transverse diameter is 17cm, the plant is medium-maturing, and the plant is planted for about 80 days after harvesting.
Disease resistance is two major diseases of broccoli under field natural disease conditions: the incidence rates of cauliflower virus disease and cauliflower black rot are investigated and counted.
The analysis of the nutritional quality mainly comprises the contents of water, vitamin C (Vc), soluble sugar, titratable acid, soluble solid, crude fiber, protein, mineral, anthocyanin and glucosinolate of the curd flower, and the determination method comprises the following steps:
the moisture measurement adopts a national standard method GB/T5009.3-2010; vc determination adopts a national standard method GB 6195-; the soluble sugar is measured by a national standard method GB 6194-1986; the titratable acid is determined by adopting a national standard method GB/T12456-2008; the soluble solid substance is measured by a refractometer method; the crude fiber is measured by adopting a national standard method GB/T5009.10-2010; the protein is measured by a national standard method GB/T5009.5-2010; the mineral substance is measured by adopting a plasma emission spectrum ICP method; the anthocyanin is measured by a differential colorimetry; the thioglucoside is determined by HPLC analysis.
The combining ability measuring method comprises: the general fitness of a parent is determined by referring to the general fitness determination method described in vegetable breeders, second edition (1990), i.e., F1 prepared from the parent and other parents is used to calculate the difference between the average yield (or other trait value) and the total average of all F1 in the test.
2. The loose type cauliflower homozygous line S5 is taken as a male parent, and is hybridized with white cauliflower cytoplasmic male sterile variety Jing Ming 50 (Jing Ming Yinong (Beijing) seed industry science and technology Co., Ltd.) to obtain the first filial generation F1. The first filial generation pollen aborts, and is used as a female parent for further backcross transformation.
3. And (3) carrying out backcross transformation by taking the first filial generation F1 as a female parent and S5 as a recurrent parent, and carrying out five-generation backcross to obtain a loose cauliflower cytoplasmic male sterile line CMSS5 (white cauliflower 2). The main morphological characteristics of the CMSS5 plant and the curd are the same as S5, except that fertility is different from S5.
The screening criteria in the backcrossing process were: in the backcross process of each generation, the plants and the single plants with the flower ball shape similar to the recurrent parent S5 are selected as the female parent plants of the current generation to carry out backcross pollination with the recurrent parent S5.
The main characteristic characteristics of the loose cauliflower cytoplasmic male sterile line CMSS5 are as follows: the sterility is stable, the sterile plant rate and the sterility degree of the sterile plant are both 100%, the honey gland is developed, the nectar is more, the stigma is normal, the flowering and fruiting properties are good, and the seedlings are not yellowed under the low-temperature condition. The plant is semi-upright, the plant height is 50cm, the development degree is 55 multiplied by 60cm, 15 outer leaves are adopted, the plant is oval, light green, the leaf surface wax powder is medium, the flower ball is flat, round, loose and white, the flower stalk is 14cm long, the weight of a single ball is 0.7Kg, the ball height is 13cm, the ball transverse diameter is 17cm, the plant is medium-maturing, and the plant is planted for about 80 days after harvesting.
4. Purple compact cauliflower variety amethyst (Henan New county and pasture fine breed research institute) is cultured by using free microspore to obtain multiple regeneration homozygous plants with different characters, 4 excellent single plants with strong growth vigor, medium plant type, good disease resistance and purple flower balls are screened out through field observation and identification, and the single plants are respectively selfed to obtain 4 purple cauliflower homozygous strains (1301, 1309, 1330 and 1341). Wherein 1301 the plants are upright, the plant height is 65cm, the development degree is 58 multiplied by 60cm, 18 outer leaves are provided, the plants are oval, dark green, more leaf surface wax powder, compact flower balls, purple, 5cm long pedicel, 0.4Kg of single ball weight, late maturity and are planted for about 100 days after harvest; 1309 plants are upright, the height of the plants is 65cm, the spread is 50X 52cm, 17 outer leaves, long oval, dark green, more leaf surface wax powder, compact flower ball, purple, the flower stalk is 6cm long, the weight of a single ball is 0.6Kg, the plant is late-maturing, and the plant is planted and harvested for about 90 days; 1330 plants are semi-upright, the plant height is 62cm, the development degree is 60 multiplied by 68cm, 16 outer leaves are in long oval shape, dark green, in leaf surface wax powder, the flower ball is compact, purple, the flower stalk is 6cm long, the weight of a single ball is 0.5Kg, the medium-maturing is carried out, and the planting is carried out for about 80 days after the harvest; 1341 plants are relatively flat, 55cm high, 65X 68cm wide, 17 leaves, long oval, dark green, more wax powder on leaf surface, compact flower ball, purple, 6cm long pedicel, 0.4Kg of single ball weight, medium early maturity, and about 70 days after planting and harvesting.
The method for identifying disease resistance and analyzing nutritional quality is the same as the step 1.
5. The loose cauliflower cytoplasmic male sterile line CMSS5 is used as a female parent, and 4 purple cauliflower homozygous lines (1301, 1309, 1330 and 1341) are respectively used as male parents for hybridization to obtain F1.
6. F1 of the step 5 is subjected to a field product ratio test, and loose cauliflower Qingnong 80 and Qingnong 90 which are cultivated in a large area in production are used as control varieties. Through determination and evaluation of main agronomic characters, yield, quality, disease resistance and nutrient components, a purple loose cauliflower hybridization combination CMSS5 multiplied 1309 with excellent comprehensive characters and the serial number ZS1 is screened out, namely the nutrient-rich target loose cauliflower created by the invention.
Second, main characteristic features
1. Major agronomic traits
ZS 1: the first generation hybrid of purple loose cauliflower prepared by taking a sterile line CMSS5 as a female parent and taking a selfing line 1309 as a male parent is expressed as middle and late maturity and is harvested about 90 days after field planting; the plant type is upright, the plant height is 65cm, the spreading degree is 55 multiplied by 55cm, 17 outer leaves are adopted, the plant is grayish green, and the leaf surface has more wax powder; the flower bulbs are flat, spherical, loose and purple, the flower stalks are about 13cm long, the weight of a single ball is 1.2Kg, the height of the ball is 13cm, the transverse diameter of the ball is 21cm, the flower bulbs are ripe in middle and late periods, and the planting is carried out for about 90 days after harvesting.
2. Main nutrient components and content
The purple loose cauliflower ZS1 has the outstanding characteristics in the aspect of nutrition and health care value that: has anthocyanin which is not possessed by common white loose cauliflower, the average content of the anthocyanin of the ZS1 is 42.30mg/100gFW, and the anthocyanin is not detected in the curd of the control variety Qingnong 90.
The results of the measurement of the main nutrient components and contents of the ZS1 are shown in tables 1 to 3, and the Qingnong 90, a main white loose cauliflower variety with consistent maturity, was used as a Control (CK).
TABLE 1 determination of essential nutrient composition of nutrient-rich loose cauliflower ZS1
In table 1, "%" in the rows "ZS 1" and "qingnong 90 (CK)" both represent mass percentages, "plus or minus% vs control CK represents the percentage change of each nutrient component of ZS1 from the control,"/"represents no anthocyanin.
TABLE 2 determination of essential mineral nutrient content of nutrient-rich loose cauliflower ZS1
Unit: mg/100gFW
In table 2, "CK + -%" compared to control indicates the percentage change of various mineral nutrients of ZS1 compared to control.
TABLE 3 determination of the major glucosinolate component of nutrient-rich bulk cauliflower ZS1
Unit: mu mol/gFW
In table 3, "CK + -%" versus control indicates the percent change of each glucosinolate component of ZS1 versus control.
Compared with the common loose cauliflower Qingnong 90, the nutrient-rich loose cauliflower ZS1 has the advantages that Vc, titratable acid, soluble solid, crude fiber and protein are all remarkably improved, and are respectively improved by 39.03%, 56.25%, 7.02%, 2.94% and 24.43%; the main mineral nutrients are also obviously improved, and the calcium, potassium, magnesium, sodium, phosphorus, copper, iron, manganese and zinc are respectively improved by 22.55%, 15.53%, 13.63%, 22.61%, 13.86%, 16.47%, 54.12%, 5.71% and 12.14%; the main glucosinolate components are also remarkably improved, and the IBE, SIN, RAA, NAP, 4OH, GBC, 4ME and NEO are respectively improved by 148.15%, 185.71%, 50.00%, 5.26%, 40.00%, 92.29%,
12.86% and 1450.00%, only PRO content 61.29% lower than the control, and there have been scientific studies that show that PRO is a precursor of goitrogen 5-etazolidine-2-thione (Heaney & Fenwick, 1995) and that lower levels are more beneficial to human health.
Thirdly, the technical points of propagation and seed production
1. Parent propagation: adopting a sunlight greenhouse or improving a cold frame to collect seeds. In the female parent breeding shed (chamber), the sterile line and corresponding maintainer line are in a ratio of 2: 1 proportion planting. The flowering period is isolated by gauze, bees are released in the shed (room) for pollination, and the maintainer line is pulled out after the flowering period is finished. And (4) collecting seeds from the sterile line plants to obtain female parent seeds. The male parent seed-collecting shed (room) is pollinated by bees in the flowering period, and male parent seeds are obtained by collecting.
2. Hybrid first generation seed production technology:
(1) strict isolation: strict isolation measures must be taken in the seed production base, preferably natural barrier isolation and time isolation, and if space isolation is used, the isolation distance should be more than 2000 meters.
(2) Adjusting the seeding time: the white cauliflower sterile line CMSS5 bred by the method is used as a female parent, and the purple cauliflower excellent selfing line 1309 is used as a male parent for hybrid seed production. Because the flowering phases of the male parent and the female parent are different, in order to ensure that the flowering phases of the male parent and the female parent meet each other and improve the seed collection yield during seed production, the seeding phases of the male parent and the female parent need to be adjusted according to the climatic conditions of a seed production field during seeding.
(3) Planting ratio: male parent: the female parent is as follows: 2 or 1: 3, and performing permanent planting.
(4) Field management: properly squatting after the permanent planting and seedling recovering of the male parent and the female parent, and controlling water and fertilizer management. The cross-shaped ball is cut after the ball is 12-13 cm, so that bolting and flowering are promoted. Releasing bees for pollination after blooming, and checking and removing the mixed plants in time. And (5) enhancing the management of plant diseases and insect pests in the field.
(5) Seed harvesting: after the flowering phase, the parent strain is removed and the seed is harvested from the parent strain.
Claims (1)
1. The cultivation method of the nutrient-rich loose cauliflower comprises the following steps:
A1) hybridizing white cauliflowers respectively named as white cauliflower 1 and white cauliflower 2 to obtain a first filial generation; the white cauliflower 1 is a homozygous plant obtained by culturing free microspores of a loose cauliflower variety Qingnong 90;
the white cauliflower 2 is Jing Ming 50 or the descendant thereof;
A2) taking the first filial generation of A1) as a female parent, taking the white cauliflower 1 as a recurrent parent, carrying out continuous 5-generation backcross with the first filial generation, and screening to obtain a backcross progeny of cytoplasmic male sterility;
A3) hybridizing the backcross progeny serving as a female parent and the purple cauliflower serving as a male parent, and screening to obtain the nutrient-rich loose cauliflower; the purple cauliflower is a homozygous plant obtained by culturing purple crystals with free microspores;
the nutrient-rich bulk cauliflower has an increased nutritional value as compared to the white cauliflower 1;
the increase in nutritional value is reflected in the following 1) and 2):
1) the nutrient-rich bulk cauliflower anthocyanidin, vitamin C, titratable acid, soluble solids, crude fiber, protein, minerals, and glucosinolates beneficial to the human body are increased in content as compared to the white cauliflower 1; the mineral substances are calcium, potassium, magnesium, sodium, phosphorus, copper, iron, manganese and zinc; the glucosinolates beneficial to human body comprise RAA, NAP, 4OH, GBC, 4ME and NEO;
2) compared with the white cauliflower 1, the nutrient-rich loose cauliflower has reduced content of glucosinolate which is not beneficial to human body; the glucosinolate component not beneficial to human body is PRO.
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