CN106106122A - Leaf produces the breeding method of astaxanthin tomato variety - Google Patents

Abstract

Breeding method of new tomato variety 'Yunxia 2', using the old Yunnan local variety 'Dapingding' as the female parent and the astaxanthin-producing engineering tomato 'Shrimphong 1' as the male parent, and using acetone to extract the astaxanthin leaves element, and its content was detected by UPLC method. 'Yunxia 2' combines the main excellent traits of the parents, with good fruit quality and high astaxanthin production in leaves. Its main characteristics are limited growth type of plant, plant height about 130cm, dark red, robust growth; yellow-green to light brown stem, strong branching ability; ordinary leaf type, yellow-green to dark red petiole, dark red leaves, stem The average content of the substance astaxanthin is 2.27mg/g. Calyx dark red, radiant corolla orange-yellow to orange-red, anthers orange-red, stigma and persistent young fruit brown-red. The fruit is bright red, the fruit shoulder is purple, round to oblate, the fruit shape index is 0.67-0.79, the fruit surface is smooth, bright, the flesh is thick, pink, the placenta jelly is red, the ventricle is 5-7, and the hardness is moderate , the average single fruit weight is 150g, and the maximum single fruit weight can reach 350g.

Description

Breeding method of leaf-producing astaxanthin tomato varieties

Technical field:

The invention belongs to the field of biotechnology, and specifically relates to a method for cultivating tomato plant hybrid varieties, and more specifically, relates to a method for hybridizing tomato plant hybrids with 'Dapingding' as the female parent and astaxanthin-producing engineering tomato 'Xiahong No. 1' as the male parent. The invention discloses a method for cultivating a single-cross variety with good fruit quality and high yield of astaxanthin in leaves.

Background technique:

'Dapingding' is the earliest local old variety planted in Yuanmou County, Chuxiong Prefecture, Yunnan Province. Its plant is a limited growth type, with a plant height of about 120cm. It grows vigorously and has strong branching ability. The main stem has 8 to 10 leaves and bears the first inflorescence. , a mid-to-late maturing variety; strong adaptability, heat resistance, easy cultivation, and high fruit setting rate. The fruit shape is neat, flat and round, the fruit surface is evenly colored, smooth without edges, and bright in luster. The ripe fruit is bright red, with thick flesh, 5 to 7 ventricles, and moderate hardness. The average single fruit weight is 150g, and the maximum single fruit weight can reach 350g. It is cultivated in spring greenhouses in Kunming, and it takes about 160 days from sowing to harvesting. Astaxanthin-producing engineering tomato 'Xiahong No. 1' has no difference in plant, flower, leaf and fruit shape from ordinary tomato, but the difference is mainly in color. Specifically, the plant is limited growth type, and the plant height is about 110 cm. The stem is semi-erect, 2.6-4.7cm thick, with 4-6 branches, yellow-green to light brown. Common leaf shape, petiole green to dark red, leaves dark red. Hermaphrodite flowers, calyx dark red, radial corolla orange-yellow to orange-red, anthers orange-red, stigma and persistent young fruit brown-red. The fruit is nearly spherical, orange-red when ripe, with an average single fruit weight of about 100g, and the average astaxanthin content of the fruit is 1.63mg/g.

The cultivation method of a new tomato variety with excellent fruit and leaf quality is obtained by cross breeding with the old Yunnan local variety 'Dapingding' as the female parent and the astaxanthin-producing engineering tomato 'Xiahong 1' as the male parent. And technology has not been reported yet.

Invention content:

The object of the present invention is to provide a method for cultivating a new tomato variety with good fruit quality and high astaxanthin-yielding leaves by using common cultivated tomato as the female parent and astaxanthin-producing engineered tomato as the male parent for cross breeding.

Above-mentioned purpose of the present invention is achieved by following technical scheme:

The breeding method of tomato variety 'Yunxia 2', including pollen collection, female parent emasculation, artificial pollination, fruit harvesting, seed treatment, astaxanthin content determination steps, using 'Dapingding' as the female parent, producing shrimp green Astaxanthin was extracted with acetone and its content was detected by UPLC method.

As the breeding method of tomato variety 'Yunxia No. 2', the pollen collection is that the male parent is sown 20 days earlier than the female parent, and the male parent 'Xiahong No. 'For flowers with reddish petals at 170-190°, pick out the stamens, spread them out on white paper, mark the variety name and date of collection, and place them in a clean place to dry naturally in the shade. When the anthers split and the pollen is scattered, collect The pollen is reserved; the female parent is emasculated on the day before pollination by selecting the 'big flat-top' corolla elongated and exposed, the color turns from green to yellow, the petals are not open or slightly open, the anthers are yellow-green, but the pollen is not cracked, The flower buds that bloom the next day are detasseled; the artificial pollination is when the dew in the female field dries up every day, at 8-9 o'clock in the morning on a sunny day, and the temperature is 20-25°C; Harvest the mature fruit in 60 days; the seed treatment is to harvest the fully red ripe fruit, and then take the seeds after 1-2 days after ripening; the astaxanthin content determination is to extract the astaxanthin from the leaves with acetone, The content was detected by UPLC method.

Such as the cultivation method of the tomato variety 'Yunxia No. 2', wherein the pollen collected in the pollen collection step is temporarily stored in a refrigerator at 4°C, and stored in a refrigerator at -80°C with silica gel as a desiccant for long-term storage.

Such as the breeding method of the tomato variety 'Yunxia No. 2', wherein the step of detasseling the female parent is to use the left hand to hold the base of the flower bud steadily, and the right hand to clamp the anther with tweezers to remove all the stamens, or to peel off the petals together , pay attention to clean emasculation, pay attention to isolation after emasculation, and remove flowers and self-bred fruits that are not suitable for pollination.

Such as the cultivation method of the tomato variety 'Yunxia No. 2', wherein the most suitable average temperature for the artificial pollination step is 20-25°C, and a small amount of pollen is picked up by dipping the rubber tip of an ordinary pencil to the female parent. On the stigma of the male flower, or put the pollen directly on the white paper, directly contact the emasculated stigma, and dip a small amount of pollen. Use a 6×8cm tea bag to cover the bag, fix it with a paper clip, and mark it with a tag. The parent combination number and pollination date are marked on the tag. Take out the bag 10 days after pollination, observe the fruit setting at the same time, pruning and cleaning in time, and remove unpollinated flower buds and newborn ones. Inflorescences, pruning and pruning, removal of redundant axillary buds and elongated branches, coordinated management of water and fertilizer and pest control.

Such as the breeding method of the tomato variety 'Yunxia No. 2', wherein in the fruit harvesting step, only fruits with normal growth and development are harvested, and unmarked fallen fruits and deformed fruits are not harvested.

Such as the cultivation method of the tomato variety 'Yunxia No. 2', wherein in the seed treatment step, the fruit is cut horizontally with a knife, and the seeds and pulp are squeezed into an anhydrous non-metal container to ferment for 1-2 days , until the surface of the fermented product is white, stir with a wooden stick, the seeds sink, pinch the seeds by hand, the pectin is separated from the seeds, wash the seeds when there is no sticky feeling, remove the pulp, sundries and deflated seeds, and put them into double-layer gauze Squeeze the water out of the bag, spread it on canvas or absorbent paper at the bottom to dry in the shade, turn it over several times, so that the water on the surface of the seeds evaporates quickly, until the water content of the seeds drops to 7% to 8%, pack them in paper bags, and store them at room temperature. Sow in March of the following year.

Such as the breeding method of the tomato variety 'Yunxia No. 2', wherein the determination of astaxanthin is as follows: when the plant grows up, collect the 4-5 layers of leaves under the top of the new tomato variety 'Yunxia No. 2' , dry leaf powder was obtained by freeze-drying, 50 mg of sample was weighed, extracted 4 times with acetone until the extract was colorless, the extract was collected and the volume was adjusted to 5ml, and the sample was loaded to determine the pigment after suction filtration, and 3 repetitions were set. The content of astaxanthin was determined by ultra-high performance liquid chromatography.

The cultivation method of a tomato variety 'Yunxia No. 2' of the present invention comprises the following steps: (1) pollen collection, the male parent is sown 20 days earlier than the female parent, and the male parent is collected after the dew dries up in the morning during the full flowering period. Shrimp Red No. 1' petals spread 170-190° and the flower color is reddish. Use tweezers to pick out the stamens indoors, spread them on a thin and flexible white paper, mark the variety name and collection date, and put them in a clean place for natural Dry in the shade, when the anthers split and the pollen comes out, collect the pollen for later use, temporarily store the pollen in a refrigerator at 4°C, and store it in a refrigerator at -80°C with silica gel as a desiccant for long-term storage; (2) emasculate the female parent, the day before pollination Choose 'Big Flat Top' corolla elongated and exposed, the color turns from green to yellow, the petals are not opened or slightly opened, the anthers are yellow-green, but the pollen is not cracked, and the flower buds that bloom the next day are detasseled. When emasculating, the left hand Hold the base of the flower bud steadily, hold the anther with tweezers in the right hand, remove all the stamens, or peel off the petals together, isolate after emasculation, and remove flowers and self-bred fruits that are not suitable for pollination; Afterwards, pollination begins, and it is carried out at 8-9 in the morning on sunny days. The suitable average temperature for pollination is 20-25°C. Use the rubber tip of an ordinary pencil to dip a small amount of pollen, and point it on the stigma of the female parent's emasculated flower or put the pollen on it. On white paper, directly touch the emasculated stigma, dip in a small amount of pollen, cover it with a 6×8cm tea bag, fix it with a paper clip, and mark it with a tag. The parental combination number and pollination date are marked on the tag, and the bag is taken out 10 days after pollination. At the same time observe the fruit setting situation, pruning and cleaning in time, remove unpollinated flower buds and new inflorescences, pruning and branching, remove redundant axillary buds and elongated branches, and carry out coordinated management of water and fertilizer and prevention and control of diseases and insect pests; (4) fruit harvesting, 50 days after pollination Harvest mature fruits within ~60 days, only harvest fruits with normal growth and development, and do not harvest unmarked fallen fruits and deformed fruits; (5) Seed treatment, after harvesting fully red ripe fruits, post-ripen for 1 to 2 days Take the seeds again, cut the fruit horizontally with a knife, squeeze the seeds and glue into an anhydrous non-metal container and ferment for 1 to 2 days, until the surface of the fermented product is white, stir with a wooden stick, the seeds sink, and pinch the seeds with your hands , the pectin is separated from the seeds, and the seeds can be cleaned without sticky feeling, rinsed with clean water to remove the pulp, sundries and deflated seeds, put them in a double-layer gauze bag to squeeze out the water, and spread them on canvas or absorbent paper at the bottom , turn over, so that the water on the surface of the seeds evaporates quickly, until the water content of the seeds drops to 7% to 8%, they are packed in paper bags, stored at room temperature, and sown in March of the following year; (6) Determination of astaxanthin content, new tomato variety 'Yun Shrimp No. 2' is sown and waited for the plant to grow up, harvest the top 4-5 layers of leaves, freeze-dry to obtain leaf dry powder, weigh 50 mg of the sample, add acetone and extract 4 times until the extract is colorless, collect and extract The solution was adjusted to 5ml, and after suction filtration, the sample was loaded to determine the pigment, and three repetitions were set up, and the content of astaxanthin was determined by ultra-high performance liquid chromatography (UPLC).

Such as the cultivation method of the tomato variety 'Yunxia No. 2', wherein the 'Yunxia No. 2' obtained by crossing 'Dapingding' as the female parent and the astaxanthin-producing engineering tomato 'Xiahong No. 1' as the male parent The biological characteristics of Shrimp No. 2' are: "Yun Xia No. 2" combines the main excellent traits of the parents, the fruit quality is good, and the leaves produce high astaxanthin. Its main characteristics are: limited growth of plants, plant height of about 130cm , dark red, strong growth; yellow-green to light brown stems, strong branching ability; common leaf type, yellow-green to dark red petioles, dark red leaves, the average content of astaxanthin in dry matter is 2.27mg/g, calyx Dark red, radial corolla orange-yellow to orange-red, anthers orange-red, stigma and persistent young fruit brown-red, fruit bright red, fruit shoulder purple-red, round to oblate, fruit shape index 0.67-0.79, fruit Smooth surface, bright luster, thick, pink flesh, red placenta jelly, 5-7 ventricles, moderate hardness, average single fruit weight 150g, maximum single fruit weight 350g.

Compared with the prior art, the advantage of the present invention lies in that the tomato variety 'Yunxia 2' cultivated by the cultivation method of the present invention takes the average content of astaxanthin in the dry matter of the leaves as the target trait of 2mg/g, taking into account the fruit Characters, whether the plant color is dark red or not is used as a marker character for distinguishing hybrid varieties and 'big flat top'. Combining phenotype observation and determination and evaluation of astaxanthin content in leaves, 'Yunxia 2' combined the main excellent traits of the parents, with good fruit quality and high astaxanthin production in leaves. Its main characteristics are limited growth type of plant, plant height of about 130cm, dark red, strong growth; yellow-green to light brown stem, strong branching ability; ordinary leaf type, yellow-green to dark red petiole, dark red leaves, other The average content of astaxanthin in dry matter is 2.27mg/g. Calyx dark red, radiant corolla orange-yellow to orange-red, anthers orange-red, stigma and persistent young fruit brown-red. The fruit is bright red, the fruit shoulder is purple, round to oblate, the fruit shape index is 0.67-0.79, the fruit surface is smooth, bright, the flesh is thick, pink, the placenta jelly is red, the ventricle is 5-7, and the hardness is moderate , the average single fruit weight is 150g, and the maximum single fruit weight can reach 350g.

detailed description:

Further illustrate the substantive content of the present invention below with the actual example of the present invention, but do not limit the present invention with this.

Example 1:

Hybridization method of tomato variety 'Yunxia 2':

It is obtained by crossing the old Yunnan local variety 'Dapingding' as the female parent and the astaxanthin-producing engineering tomato 'Xiahong No. 1' as the male parent.

The female parent 'Dapingding' tomato is the earliest local old variety planted in Yuanmou County, Chuxiong Prefecture, Yunnan Province. The fruit surface is smooth, large in size and good in quality. . The male parent 'Xiahong 1' is an engineering tomato producing astaxanthin, which was bred in 2009. It was screened and isolated from green algae by Huang Junchao et al. (Huang JC et al., 2013) into plant cells that can catalyze The ketolase and hydroxylase genes of β-carotene and zeaxanthin to synthesize astaxanthin were transferred into wild-type tomato plants (cv. Preserved by the General Microorganism Center of the Culture Collection Management Committee, the preservation number is: CGMCC No. 9223, and the classification name is: 'Shrimp Red No. 1' transgenic tomato). From 2010 to 2012, they were planted in potted plants for 4 generations in the School of Biological Sciences of the University of Hong Kong and the Kunming Institute of Botany, Chinese Academy of Sciences, with stable traits. Sowing started in March 2013. 'Xiahong 1' was sown 20 days earlier than 'Dapingding'. The day before pollination in May, the corolla of 'Dapingding' was extended and exposed. The color changed from green to yellow, and the petals were not opened. Or slightly open, the anthers are yellow-green, but the pollen is not cracked, and the flower buds that bloom the next day are emasculated, pay attention to isolation. When the dew dries up in the morning, collect the male parent 'Xiahong No. 1' whose petals spread 170-190° and the flower color is reddish. Pick out the stamens indoors with tweezers, spread them out and dry them on a thin and flexible white paper, and mark the collected flowers. date, put it in a clean place to dry naturally. When the anthers are dehiscated and the pollen is scattered, collect the pollen for later use. The next day, after the dew in the mother field dries up, pollination is carried out at 8-9 am on a sunny day. Specifically, use the rubber tip of an ordinary pencil to pick up a small amount of pollen and instill it on the stigma of the female parent's emasculated flower, or put the pollen on a white paper, touch the emasculated stigma, and directly dip in a small amount of pollen. Cover the bags with 6×8cm tea bags, fix them with paper clips, put them on a tag, and mark the combination number of the parents and the date of pollination on the tag. Take out the bag 10 days after pollination, observe the fruit setting at the same time, pruning and cleaning in time, remove unpollinated flower buds and new inflorescences, pruning and branching, and remove excess axillary buds and elongated branches. During the period, pay attention to the coordinated management of water and fertilizer and the prevention and control of diseases and insect pests. Ripe fruits can be harvested 50-60 days after pollination. Unmarked fallen fruits and deformed fruits are not harvested, only fruits with normal growth and development are harvested. After the fully red ripe fruit is harvested, it can be ripened for 1 to 2 days before replanting. When picking the seeds, cut the fruit horizontally with a knife, then squeeze the seeds and pulp into an anhydrous non-metallic container for fermentation, wash the seeds after 1-2 days of fermentation, put them in a ventilated and dry place to filter the water and dry them immediately , Store until the moisture content of the seeds drops to 7% to 8%, and sow in March of the following year. After the tomato variety 'Yunxia No. 2' grows up, the 4-5 layers of leaves on the top and bottom are harvested in good time, and freeze-dried to obtain dry powder. Accurately weigh 50 mg of the sample, add acetone and extract 4 times until the extract is colorless, collect the extract and dilute it to 5ml, apply the sample to determine the pigment after suction filtration, set 3 repetitions, and use ultra-high performance liquid chromatography (UPLC ) method to determine its astaxanthin content. After 4 times of planting in 2014 and 2015, the results of phenotype observation and determination of astaxanthin content in leaves showed that 'Yunxia 2' has good and neat traits, good fruit quality, high content of astaxanthin in leaves, and excellent disease resistance. powerful.

The traits of the hybrid parents involved in the hybridization method of the present invention are as follows:

'Dapingding': the earliest local old variety planted in Yuanmou County, Chuxiong Prefecture, Yunnan Province. The plant is a limited growth type, with a plant height of about 120cm, strong growth, and the first inflorescence on the main stem with 8-10 leaves. It is a mid-late maturing variety; Strong adaptability, heat resistance, easy cultivation, high fruit setting rate. The fruit shape is neat, flat and round, the fruit surface is evenly colored, smooth without edges, and bright in luster. The ripe fruit is bright red, with thick flesh, 5 to 7 ventricles, and moderate hardness. The average single fruit weight is 150g, and the maximum single fruit weight can reach 350g. It is cultivated in spring greenhouses in Kunming, and it takes about 160 days from sowing to harvesting.

Astaxanthin-producing engineering tomato 'Xiahong No. 1': its plants, flowers, leaves, and fruits are no different from ordinary tomatoes in shape, but the difference is mainly in color. Specifically, the plant is limited growth type, and the plant height is about 110 cm. The stem is semi-erect, 2.6-4.7cm thick, with 4-6 branches, yellow-green to light brown. Common leaf shape, petiole green to dark red, leaves dark red. Hermaphrodite flowers, calyx dark red, radial corolla orange-yellow to orange-red, anthers orange-red, stigma and persistent young fruit brown-red. The fruit is nearly spherical, orange-red when ripe, with an average single fruit weight of about 100g, and the average astaxanthin content of the fruit is 1.63mg/g.

The new tomato variety 'Yunxia 2' combines the main excellent traits of the parents, with good fruit quality and high yield of astaxanthin in leaves. Its main characteristics are limited growth type of plant, plant height of about 130cm, dark red, strong growth; yellow-green to light brown stem, strong branching ability; ordinary leaf type, yellow-green to dark red petiole, dark red leaves, other The average content of astaxanthin in dry matter is 2.27mg/g. Calyx dark red, radiant corolla orange-yellow to orange-red, anthers orange-red, stigma and persistent young fruit brown-red. The fruit is bright red, the fruit shoulder is purple, round to oblate, the fruit shape index is 0.67-0.79, the fruit surface is smooth, bright, the flesh is thick, pink, the placenta jelly is red, the ventricle is 5-7, and the hardness is moderate , the average single fruit weight is 150g, and the maximum single fruit weight can reach 350g.

Claims (10)

1. the breeding method of tomato variety ' cloud shrimp No. 2 ', including pollen collection, maternal emasculation, artificial pollination, fruit harvesting, kind Son processes, content astaxanthin determination step, and with ' big flat-top ' for maternal, producing astaxanthin engineering Fructus Lycopersici esculenti ' red No. 1 of shrimp ' is that male parent is entered ' cloud shrimp No. 2 ' is cultivated in row hybridization, with acetone extraction blade astaxanthin, and uses UPLC method to detect its content.

2. the breeding method of tomato variety ' cloud shrimp No. 2 ' as claimed in claim 1, it is characterised in that: described pollen collection It is that male parent gathers male parent ' red No. 1 of shrimp ' petal and launches 170-after dew is dry than maternal planting early enough 20 days, full-bloom stage the morning 190 ° and the reddest flower of pattern, choose stamen, spread out on blank sheet of paper, indicates kind name and gathers the date, being placed in clean place Naturally dry in the shade, treat Anther dehiscence, when pollen sheds, collect pollen standby；Described maternal emasculation is to select the previous day in pollination The elongation of ' big flat-top ' corolla exposes, and color from green to yellow, do not open or crack by petal, and flower pesticide is yellow green, but the flower that sheds that do not ftractures Powder, the alabastrum bloomed next day carries out emasculation；Described artificial pollination is after maternal field dew is done, fine day 8-9 in morning point, temperature 20～25 DEG C of pollinations；Described fruit harvesting is 50～60 days mature fruits of gathering after pollination；Described seed treatment has been After complete red ripe fruit harvesting, treat after-ripening 1～take kind again in 2n days；It is with acetone extraction blade shrimp that described content astaxanthin measures Blue or green element, UPLC method detects its content.

3. the breeding method of tomato variety ' cloud shrimp No. 2 ' as claimed in claim 2, it is characterised in that: described pollen collection The pollen collected in step is temporarily stored with 4 DEG C of refrigerator cold-storages, and long-term storage silica gel is stored in-80 DEG C of ice as desiccant Case.

4. the breeding method of tomato variety ' cloud shrimp No. 2 ' as claimed in claim 2, it is characterised in that: described maternal emasculation Step is surely to hold alabastrum base portion with left hand, and right hand tweezers are clamped flower pesticide and all extractd by its stamen, or together shell together with petal Falling, emasculation is clean, isolates after emasculation, rejects flower and the selfing fruit being not suitable for pollination simultaneously.

5. the breeding method of tomato variety ' cloud shrimp No. 2 ' as claimed in claim 2, it is characterised in that: described artificial pollination Step temperature is 20～25 DEG C, dips a small amount of pollen point by the rubber tip of common pencil and invests on the stigma of maternal emasculation flower, Or directly pollen is placed on blank sheet of paper, directly contacts the stigma of emasculation, dip a small amount of pollen.With 6 × 8cm tea bag bag bagging, Paper clip is fixed, listing mark, and plate is indicated Parent combination number and pollination date, and bag taking after pollinating 10 days is seen simultaneously Examine the situation of bearing fruit, cleaning of pruning in time, remove the alabastrum and the inflorescence of new life do not pollinated, training is pruned, and extracts unnecessary axil Bud and the branch of excessive growth, carry out liquid manure coordinated management and the prevention and control of plant diseases, pest control.

6. the breeding method of tomato variety ' cloud shrimp No. 2 ' as claimed in claim 2, it is characterised in that: described fruit harvesting Step is only gathered the normal fruit of growth promoter, do not gather unmarked fallen fruits and malformed fruit.

7. the breeding method of tomato variety ' cloud shrimp No. 2 ' as claimed in claim 2, it is characterised in that: described seed treatment Fruit is laterally cut open with knife by step, ferments 1～2 day in seed and glue flesh are clamp-oned anhydrous nonmetallic vessel, to fermentation Thing surface is white, stirs with wooden stick, and seed sinks, and pinches seed with hands, and pectin separates with seed, cleans and plant during the glutinous sliding sense of nothing Son, removes sarcocarp, foreign material and flat seed, squeezes the water out in loading double gauze bag, is spread out in the built on stilts canvas in bottom or absorbent paper Dry in the shade, stir several times, make the moisture of the surface of the seed evaporate rapidly, during until seed moisture content is down to 7%～8%, paper bag bag Dress, room temperature storage, sow March in next year.

8. the breeding method of tomato variety ' cloud shrimp No. 2 ' as claimed in claim 2, it is characterised in that: described astaxanthin Determination step is, plant to be planted is grown up, and gathers the lower 4-5 floor blade in New Tomato Variety ' cloud shrimp No. 2 ' top, freeze-dried acquisition blade Dry powder, weighs sample 50mg, adds acetone extraction 4 times, until extract is colourless, collects extract and is settled to 5ml, loading after sucking filtration Measure pigment, 3 repetitions are set, use ultra-performance liquid chromatography to measure the content of its astaxanthin.

9. the breeding method of tomato variety ' cloud shrimp No. 2 ', it is characterised in that it comprises the steps: (1) pollen collection, male parent ratio Maternal planting early enough 20 days, full-bloom stage gathers male parent ' red No. 1 of shrimp ' petal expansion 170-190 ° and pattern is inclined after dew is dry the morning Red flower, chooses stamen in indoor tweezers, spread out thin and on the blank sheet of paper of flexible, indicate kind name and gather the date, put Naturally drying in the shade in clean place, treat Anther dehiscence, when pollen sheds, collect pollen standby, pollen is stored temporarily in 4 DEG C of ice Case, long-term storage silica gel is stored in-80 DEG C of refrigerators as desiccant；(2) maternal emasculation, pollination selects ' big flat-top ' the previous day Corolla elongation exposes, and color from green to yellow, do not open or crack by petal, and flower pesticide is yellow green, but the pollen that sheds that do not ftractures, next day The alabastrum bloomed carries out emasculation, and during emasculation, left hand surely holds alabastrum base portion, and right hand tweezers are clamped flower pesticide and all plucked by its stamen Remove, or together peel off together with petal, isolate after emasculation, reject flower and the selfing fruit being not suitable for pollination simultaneously；(3) manually award Powder, starts pollination after maternal field dew is dry, fine day the morning 8-9 point carry out, suitable temperature on average of pollinating is 20～25 DEG C, use The rubber tip of common pencil dips a small amount of pollen, and point is invested on the stigma of maternal emasculation flower or is placed on blank sheet of paper by pollen, Directly contacting the stigma of emasculation, dip a small amount of pollen, with 6 × 8cm tea bag bag bagging, paper clip is fixed, and listing mark, on plate Indicate Parent combination number and pollination date, bag taking after pollinating 10 days, observe situation of bearing fruit, cleaning of pruning in time simultaneously, go Except the alabastrum do not pollinated and the inflorescence of new life, training is pruned, and extracts unnecessary axillalry bud and the branch of excessive growth, carries out liquid manure coordinated management Preventing and treating with pest and disease damage；(4) fruit harvesting, 50～60 days mature fruits of gathering after pollination, the normal fruit of growth promoter of only gathering Real, do not gather unmarked fallen fruits and malformed fruit；(5) seed treatment, after complete red ripe fruit harvesting, after-ripening 1～2 days are again Take kind, laterally fruit is cut open with knife, ferment 1～2 day in seed and glue flesh are clamp-oned anhydrous nonmetallic vessel, until fermentation Thing superficial white, stirs with wooden stick, and seed sinks, and pinches seed with hands, and pectin separates with seed, without glutinous sliding sense kind the most capable of washing Son, rinses with clear water, removes sarcocarp, foreign material and flat seed, squeezes the water out in loading double gauze bag, is spread out in the canvas that bottom is built on stilts Or in absorbent paper, stir, make the moisture of the surface of the seed evaporate rapidly, during until seed moisture content is down to 7%～8%, paper bag Packaging, room temperature storage, sow March in next year；(6) content astaxanthin measures, and New Tomato Variety ' cloud shrimp No. 2 ' after planting plant to be planted is long Greatly, the lower 4-5 layer blade in top of gathering, freeze-dried acquisition blade dry powder, weigh sample 50mg, add acetone extraction 4 times, until taking out Extract is colourless, collects extract and is settled to 5ml, and after sucking filtration, loading measures pigment, arranges 3 repetitions, uses ultra high efficiency liquid phase color Spectrum UPLC method measures the content of its astaxanthin.

10. the breeding method of the tomato variety ' cloud shrimp No. 2 ' as described in claim 1 or 9, it is characterised in that described by ' big Flat-top ' it is maternal, producing astaxanthin engineering Fructus Lycopersici esculenti ' red No. 1 of shrimp ' is the biological characteristics that male parent carries out hybridizing ' the cloud shrimp No. 2 ' that obtain Property is: ' cloud shrimp No. 2 ' is combined with the main merit of Parent, and fruit quality is good, and blade high-yield astaxanthin, and it is the most special Levy as plant determinate growth, plant height 170-190cm, kermesinus, robust growth；Stem yellow green is the most light brown, branch ability pole By force；Common blade profile, petiole yellow green to kermesinus, blade kermesinus, its dry astaxanthin average content reaches 2.27mg/g, flower Calyx kermesinus, radial corolla is orange-yellow, and to Chinese red, flower pesticide is orange to red, stigma and harbor young fruit brownish red, and fruit is big Redness, fruit shoulder aubergine, circular to oblate, fruit shape index 0.67～0.79, fruit surface smooth, gloss is bright, and sarcocarp is thick, pink colour, Placenta jelly is red, ventricle 5～7, and hardness is moderate, mean fruit weight 150g, and maximum single fruit weighs 350g.