CN103392586A - Breeding method of drought resistant and cold tolerant Saccharum varieties/lines containing Erianthus fulvus Ness consanguinity - Google Patents
Breeding method of drought resistant and cold tolerant Saccharum varieties/lines containing Erianthus fulvus Ness consanguinity Download PDFInfo
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Abstract
The invention discloses a breeding method of drought resistant and cold tolerant Saccharum varieties/lines containing Erianthus fulvus Ness consanguinity. The method provided in the invention makes use of Erianthus fulvus Ness wild species (E. fulvus) and Saccharum cultivars (Saccharum spp.) to perform "interspecific distant hybridization". The hybrid progenies undergo selection from a seedling stage, i.e. under conditions (dryland, high altitude) with natural environmental stress, thus obtaining new drought resistant and cold tolerant Saccharum varieties/lines suitable to plant at an altitude of over 1300 meters.
Description
Technical field
The invention belongs to biological technical field, in particular to the breeding method of the cold-resistant sugar cane breed of a kind of drought resisting in the low latitude plateau area/be.
Background technology
Sugarcane is the most important sugar [yielding of the world and China.The development of sugarcane production, kind are crucial.Each cane planting country of the world is all by utilizing germ plasm resource and take sexual hybridization as main method, formulating new germ plasm and seed selection new varieties.20th century first half page, breeding men are by utilizing germ plasm resource " interspecific cross " and " Nobilization " in saccharum (Saccharum), and the kind of cultivating makes the cane planting zone constantly expand to the nonirrigated farmland, hills from the torrid zone to subtropics and ,Cong paddy field, temperate zone; Then 50 years because sugarcane inbreeding coefficient is large, wild germplasm reduced by generation, finally cause the sugar cane breed adaptability that is bred as wide, yield poorly, of poor benefits.So, discover and use new germ plasm resource, cultivate that breakthrough new variety of sugarcane is arranged has been imperative.On the other hand, because the cane breeding place of countries in the world is all in the torrid zone or the subtropical zone of the low height above sea level of low latitudes basically, so the sugar cane breed that its seed selection becomes exists larger limitation on planting area.The production that the sugarcane district is vast, ecotype is complicated in this and China (third place in the world Da Chantang state), particularly Yunnan Province (the second largest sugar that produces of China is economized) is actual and be mainly incompatible by introducing the present situation that adventive plants for a long time.District moves westwards along with China's sugarcane, and it is bad that the ecological condition of cane planting becomes more.So, no matter be for meeting the Competitive Needs of current Sugarcane Industry self, or, for adapting to the development trend of future source of energy sugarcane, all sugar cane breed had higher requirement.Therefore, cane breeding should be started with from enlarging genetic diversity, integrated application new technology, new method, and initiative has the new germ plasm that breeding is worth and the seed selection annidation is wide, the new variety of sugarcane of strong stress resistance, the high sugar of high yield.
More than 100 year history of sugarcane sexual hybridization breeding is in fact the process to the sugarcane germplasm development and utilization.Breeding man utilizes 2～5 kinds (species) in saccharum (Saccharum) to carry out " interspecific cross " and " Nobilization ", has created the front semicentennial splendidness of cane breeding history.But regrettably, in rear semicentennial sugarcane crossbreeding, due to arduousness and the chronicity of this work, cause many breedings man for eager for instant success and quick profits thought, mostly adopt " intervarietal cross ".Particularly China, although successively be bred as over one hundred of Sugarcane Superior Variety, brought into play due effect on producing, and they all just have the blood relationship of a few common parent, and inbreeding causes hereditary basis narrow, and what conform is indifferent.
Inbreeding must be lost genetic heterogeneity, causes vitality, adaptability, perennial root and the resistance of sugar cane breed to descend.So, need to develop new wild germplasm resources, to widen the Sugarcane genetic basis, thereby select breakthrough new variety of sugarcane.
Ravenna grass (Erianthus fulvus Ness) is the wild species (wild species) in grass family (Gramineae), Saccharinae (Saccharinae), Plumegrass (Erianthus), its drought resisting, cold-resistant, the high good characteristic of hammer degree (Brix) are that other Basic sugarcane germplasm can not reach, and in cane breeding, have value.
Summary of the invention
The present invention relates to the breeding method of the cold-resistant sugar cane breed of a kind of drought resisting that contains the Ravenna grass blood relationship/be, it comprises the steps:
(1) collection of Ravenna grass pollen and storage
In the Ravenna grass full-bloom stage, clip band blade individual plant ear stem, be placed in the well-lighted indoor clear water that carries out and supported stem, in sunny 7:00～9:00 in the morning, adopts powder to mid-September mid-August, and-80 ℃ of refrigerator storages are stand-by by after the pollen drying, being placed in;
(2) pollination hybridization
11～December, will be through the Ravenna grass pollen of low temperature storage from refrigerator, taking out, put into the refrigerator of-20 ℃, 1h. at room temperature thaws, then to preprepared sugarcane flower of female parent fringe pollination, pollinate every day 1 time, for three days on end, namely complete the pollination crossover process, after seed maturity, collect the true seed of hybrid combination;
(3) cultivate seedling
First stage is the seed sprouting stage, in the culturing room of constant temperature and humidity, carry out, ready sandy loam in small, broken bits is placed in sterilization 2～3h in high-pressure sterilizing pot, after cooling, pack into the seed tray of tiny hole, then evenly be seeded in seed in dish, with manual sprayer, spray more closely water for the first time, make seed be adjacent to native face, last earthing, just to cover seed, be advisable, once, after abundant moistening seed and soil, epiphragma is placed under 28～30 ℃ of temperature, natural scattering light and cultivates and germinate water spray again; Second stage is that seedling is heeled in the stage, and the seedling in seed tray moves on on the seedbed in plastic tunnel and carries out while growing to 4～6 leaves, heat and moisture preserving and use azophoska and survive to keep a full stand of seedings; Phase III is the Seeding planting stage, be transplanted to field heeling in when seedling grows to 40～50cm, Routine Management under natural conditions, guarantee the seedling normal growth, therefrom according to seedling growing way, damage by disease and insect, water crack, bloom or booting, hollow/Pu heart and hammer degree screen individual plant and carry out follow-up cultivation.
In a preferred embodiment of the present invention, adopt the mode of powder for shaking gently the flower fringe, with clean smooth white magazine paper, collect pollen, after removing impurity, transfer to rapidly in the culture dish of clean dried, shakeout, be positioned in the drier that silica gel is housed dry 1h under 10 ℃ of conditions.
In another preferred embodiment of the present invention, described pollination hybridization is to carry out in Hainan, and described cultivation seedling is to carry out in Kunming.
In another preferred embodiment of the present invention, described Ravenna grass is the Kunming Ravenna grass, and described sugarcane is city, precipice 89-9.
The present invention utilizes Ravenna grass wild species (E.fulvus) and cultivation of sugar cane kind (Saccharum spp.) to carry out " inter-genera distant hybridization ", its hybrid generation is namely having (nonirrigated farmland, high height above sea level) under the condition of natural environmental stress to select from the seedling stage, to obtain the cold-resistant new variety of sugarcane of suitable drought resisting in height above sea level nonirrigated farmland plantation more than 1300 meters/be.
Embodiment
Embodiment 1:
(1) collection of Ravenna grass pollen and storage
In the Ravenna grass full-bloom stage, its individual plant ear stem of clip (band blade) was placed in the well-lighted indoor clear water that carries out and supported stem, in sunny 7:00～9:00 in the morning, adopts powder to mid-September in mid-August, 2000.Fringe is spent in shake gently, with the white magazine paper that cleaning is smooth, collect pollen, after the impurity such as removal anther wall, transfer to rapidly in the culture dish of clean dried, shakeout, be positioned in the drier that silica gel is housed, dry 1h under 10 ℃ of conditions, then change in the 1.5mL plastic centrifuge tube of clean dried, be placed in-80 ℃ of refrigerators and preserve stand-by.
(2) pollination hybridization
In 11～December, will be placed in curling stone through the Ravenna grass pollen of low temperature storage from refrigerator, taking out, and from Kunming, be carried to by air, by plane cane breeding field, Hainan (Sanya Ya Cheng) same day, put into immediately the refrigerator of-20 ℃.8:00 next day takes out left and right, and then the 1h. that at room temperature thaws to maternal (city, precipice 89-9) Hua Sui pollination of preprepared sugarcane, pollinates 1 time every day, for three days on end, namely completes the pollination crossover process.After seed maturity, cut Hua Sui, the freshness protection package of packing into after the vacuum dehumidifying drying, post go back to sugarcane research institute of Yunnan Prov Agriculture University after sealing; Adopt different kinds of molecules mark (RAPD, ITS, AFLP) and in situ hybridization (GISH) technology to identify Hybrid, be about 60%～65% Hybrid and contain the Ravenna grass blood relationship.
(3) cultivation of seedling
Calendar year 2001, early March received the true seed of " city, the precipice 89-9 * Kunming Ravenna grass " hybrid combination of mailing from cane breeding field, Hainan, in sugarcane research institute of Yunnan Prov Agriculture University, carried out the cultivation of seedling immediately.divide three phases: the first stage is the seed sprouting stage, in the culturing room of constant temperature and humidity, carry out, at first by ready sandy loam in small, broken bits (sand: soil=1: 3) be placed in sterilization 2～3h in high-pressure sterilizing pot, after cooling, pack into the seed tray of tiny hole, then seed evenly is seeded in (every dish sowing 1g seed) in dish, with manual sprayer, spray more closely water for the first time, make seed be adjacent to native face, last earthing, just to cover seed, be advisable, water spray once again, after abundant moistening seed and soil, epiphragma is placed in 28～30 ℃ of temperature, under natural scattering light, cultivate and germinate, look the moisturizing of concrete condition water spray therebetween, laxative mildew-resistant and thin executing " fertile from breast ", second stage is that seedling is heeled in the stage, and the seedling in seed tray moves on on the seedbed in plastic tunnel while growing to 4～6 leaves and carry out that (heel in specification 10 * 10cm), emphasis is heat and moisture preserving and suitably uses azophoska and survive to keep a full stand of seedings, phase III is Seeding planting stage (hybrid nursery), is transplanted to field (plantation specification 80 * 50cm), Routine Management under natural conditions, guarantee the seedling normal growth heeling in when seedling grows to 40～50cm.
(4) selection of hybrid nursery
The seedling that is colonizated in field is generally estimated the combination growing way in September, selects for the first time by the end of October.Observe seedling growing way, damage by disease and insect, water crack, bloom or booting, hollow or the Pu heart, hammer degree etc., superseded growing way is poor, damage by disease and insect is many, water crack is serious, bloom or booting is severe, hollow or the Pu heart greater than 3/10, individual plant that the hammer degree is low.Select January next year for the second time, and complete observation has or not leakage choosing the lower individual plant of tracking and measuring primary election hammer degree.Selected individual plant is through following drought resisting and cold-resistant test, and approximately the hybrid generation more than 50% has better drought resisting cold resistance than maternal sugarcane.
(5) drought resistance test
Identification of Drought (is slightly coerced: blade relative water content reduction<10% by the criteria for classifying of Hsiao (1973), moderate is coerced: the reduction of blade relative water content reaches 10%～20%, severe water stress: the blade relative water content reduces>20%), when 4～5 true leaves of seedling, start to control moisture (three repetitions, take the plant of normal water supply as contrast), when plant reaches moderate and coerces, start to observe the withered situation of green grass or young crops of plant, amount of crimp and the plant height of measuring each processing, leaf area, root/shoot ratio, blade is done ratio, in leaf, arteries and veins is done the morphological indexs such as ratio, and measure plant+moisture of 1 blade, improving activity of root system, membrane permeability (electrolyte leakage rate), soluble sugar, the physiological and biochemical indexs such as malonaldehyde (MDA) and Proline (Fpro.) content, utilize in fuzzy mathematics the method for asking membership function to carry out overall merit.When plant reaches severe water stress, start rehydration and process, observation, respectively for the growing state after examination material rehydration, compared with the control, is calculated plant height injury rate.
Membership function value calculates by following formula.
In above-listed formula,
The drought resisting membership function value of expression i genotype j index, X
IjThe drought-enduring coefficient of expression i genotype j index, Xmax, Xmin represent respectively the drought-enduring coefficient that each genotype middle finger target is minimum and maximum;
For the genotypic drought resisting membership function of i average, n is the index number.Because irreducible water, rate-of-loss of coolant, soluble sugar, Proline and drought resistance are proportionate, their drought resistance membership function value calculates with the A formula; Water content, improving activity of root system, root/shoot ratio, plant height injury rate, cell leakage, malonaldehyde and drought resistance are negative correlation, so calculate with the B formula.The Physiology and biochemistry measurement result is changed into to membership function value by above-mentioned formula, with Subordinate Function, the drought-enduring coefficient of each kind is expanded on [0,1] closed interval, press the value criteria for classifying of calculating.
The mensuration of each physiological and biochemical index is all with reference to the method for Zhang Xianzheng (1992), and calculates as follows:
Leafiness rate (%)=leafiness number/blade amt * 100%
Blade is done ratio=leaf dry weight/leaf area (unit: mg/cm
2)
In leaf, arteries and veins is done arteries and veins dry weight/leaf area (unit: mg/cm in ratio=leaf
2)
Blade rate-of-loss of coolant (%)=(bright leaf weight-dehydration leaf weight)/(bright leaf weight-cured leaf weight) * 100%
Plant height is coerced index=processing plant height/contrast plant height
Plant height injury rate (%)=(contrast plant height-processing plant height)/contrast plant height * 100%
Drought-enduring coefficient 1 (%)=(arid processing costs-control treatment value)/control treatment value * 100% (for the A formula)
Drought-enduring coefficient 2 (%)=(control treatment value-arid processing costs)/control treatment value * 100% (for the B formula)
Qualification result in Table 1, table 2.As can be seen from Table 1,5 forms for the examination material, physiology, biochemical each drought resistant index have obvious difference, by the comprehensive judge of average membership function value, their strong drought resistances are weak puts in order as Yunnan sugarcane 01-117, Yunnan sugarcane 01-58, Yunnan sugarcane 01-64, Yunnan sugarcane 01-120, ROC16 (CK).
Yet the kind of drought resisting is not to be decided by its continued growth when arid, but is that it can recover later growth rapidly in arid.As can be seen from Table 5, under Water Stress Conditions, 5 plant heights for the examination material all significantly reduce, the reduction amplitude is 14.3～62.7%, wherein the plant height of ROC16 (CK) reduces the plant height of (42cm), 01-117 at most and reduces minimum (17cm), illustrates that the former drought resistance drought resistance weak, the latter is stronger; The speed that after rehydration, each material recovers to grow also has obvious difference, and it is very fast that 01-58 and 01-117 recover growth, and 01-120 and 01-64 are secondly.Result shows, the plant height performance of rehydration each material after 1 month is basically identical with their drought resisting membership function value qualification result.
The membership function value of table 1. " 89/9 * Kunming, city, precipice Ravenna grass " the drought-enduring coefficient of each index of hybrid generation
Plant height performance/cm after table 2. " 89/9 * Kunming, city, precipice Ravenna grass " hybrid generation's drought stress rehydration
(6), experiment of cold-resistance
Test is strict (DB53/T326-2010) carries out investigation and the test of each link by " sugar cane breed zoneization test specification ", the height above sea level of 5 test sites is respectively: Jing Dong---and 1350 meters, Lianghe---1630 meters, honest and just horse---1650 meters, new flat---1670 meters, Kunming---1930 meters.Result as shown in Table 3 and Table 4.
Table 3 new sugarcane lines is at the emergence rate than altitude test point (%), tillering capacity (%) and productive tiller (bar/mu) statistical form
Table 4, new sugarcane lines are in the sucrose yield than altitude test point and ranking statistical form (unit: kg/ mu)
The above, be only the specific embodiment of the present invention, but protection scope of the present invention is not limited to this, and any variation or replacement of expecting without creative work, within all should being encompassed in protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain that claims were limited.
Claims (4)
1. the breeding method of the cold-resistant sugar cane breed of the drought resisting that contains the Ravenna grass blood relationship/be, it comprises the steps:
(1) collection of Ravenna grass pollen and storage
In the Ravenna grass full-bloom stage, clip band blade individual plant ear stem, be placed in the well-lighted indoor clear water that carries out and supported stem, in sunny 7:00～9:00 in the morning, adopts powder to mid-September mid-August, and-80 ℃ of refrigerator storages are stand-by by after the pollen drying, being placed in;
(2) pollination hybridization
11～December, will be through the Ravenna grass pollen of low temperature storage from refrigerator, taking out, put into the refrigerator of-20 ℃, 1h. at room temperature thaws, then to preprepared sugarcane flower of female parent fringe pollination, pollinate every day 1 time, for three days on end, namely complete the pollination crossover process, after seed maturity, collect the true seed of hybrid combination;
(3) cultivate seedling
First stage is the seed sprouting stage, in the culturing room of constant temperature and humidity, carry out, ready sandy loam in small, broken bits is placed in sterilization 2～3h in high-pressure sterilizing pot, after cooling, pack into the seed tray of tiny hole, then evenly be seeded in seed in dish, with manual sprayer, spray more closely water for the first time, make seed be adjacent to native face, last earthing, just to cover seed, be advisable, once, after abundant moistening seed and soil, epiphragma is placed under 28～30 ℃ of temperature, natural scattering light and cultivates and germinate water spray again; Second stage is that seedling is heeled in the stage, and the seedling in seed tray moves on on the seedbed in plastic tunnel and carries out while growing to 4～6 leaves, heat and moisture preserving and use azophoska and survive to keep a full stand of seedings; Phase III is the Seeding planting stage, be transplanted to field heeling in when seedling grows to 40～50cm, Routine Management under natural conditions, guarantee the seedling normal growth, therefrom according to seedling growing way, damage by disease and insect, water crack, bloom or booting, hollow/Pu heart and hammer degree screen individual plant and carry out follow-up cultivation.
2. method according to claim 1, adopt the mode of powder for shaking gently the flower fringe, with clean smooth white magazine paper, collects pollen, after removing impurity, transfer to rapidly in the culture dish of clean dried, shakeout, be positioned in the drier that silica gel is housed dry 1h under 10 ℃ of conditions.
3. method according to claim 1, described pollination hybridization is to carry out in Hainan, described cultivation seedling is to carry out in Kunming.
4. method according to claim 1, described Ravenna grass is the Kunming Ravenna grass, described sugarcane is city, precipice 89-9.
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| CN103947540A (en) * | 2014-05-06 | 2014-07-30 | 四川省内江市农业科学院 | Breeding method for winemaking sugarcane varieties |
| CN104054484A (en) * | 2014-06-16 | 2014-09-24 | 广西南亚热带农业科学研究所 | Method for increasing survival rate of transplanted sugarcane seedlings |
| CN107494522A (en) * | 2017-08-29 | 2017-12-22 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Spontaneum pollen low-temperature storage method |
| CN108048469A (en) * | 2017-12-26 | 2018-05-18 | 云南农业大学 | The Ravenna grass class Calmodulin Gene ErCML30 expressed in Ravenna grass wild species by low temperature stress |
| CN108949780A (en) * | 2018-07-27 | 2018-12-07 | 云南农业大学 | The Ravenna grass genoid EfNAC44 that Ravenna grass wild species are expressed by low temperature stress |
| CN109197570A (en) * | 2018-10-11 | 2019-01-15 | 云南农业大学 | A method of improving sugarcane wild species resource pollen collection amount |
| CN110622811A (en) * | 2019-10-23 | 2019-12-31 | 广西壮族自治区农业科学院 | Single-plant and combined drought-resisting screening method for sugarcane seedlings |
| CN110881405A (en) * | 2019-11-27 | 2020-03-17 | 云南省农业科学院甘蔗研究所 | Method for exploring and utilizing cutting hand density breeding potential of sugarcane wild species |
| CN114009299A (en) * | 2021-12-15 | 2022-02-08 | 云南英茂糖业(集团)有限公司 | Mountain scale integrated planting method for improving per unit yield of sugarcane |
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| CN103947540B (en) * | 2014-05-06 | 2016-05-25 | 四川省内江市农业科学院 | A kind of selection of the sugar cane breed of making wine |
| CN103947540A (en) * | 2014-05-06 | 2014-07-30 | 四川省内江市农业科学院 | Breeding method for winemaking sugarcane varieties |
| CN104054484A (en) * | 2014-06-16 | 2014-09-24 | 广西南亚热带农业科学研究所 | Method for increasing survival rate of transplanted sugarcane seedlings |
| CN104054484B (en) * | 2014-06-16 | 2016-09-14 | 广西南亚热带农业科学研究所 | A kind of method improving cane seedlings transplanting success |
| CN107494522B (en) * | 2017-08-29 | 2020-05-19 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Low-temperature storage method for sugarcane pollen |
| CN107494522A (en) * | 2017-08-29 | 2017-12-22 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Spontaneum pollen low-temperature storage method |
| CN108048469A (en) * | 2017-12-26 | 2018-05-18 | 云南农业大学 | The Ravenna grass class Calmodulin Gene ErCML30 expressed in Ravenna grass wild species by low temperature stress |
| CN108949780A (en) * | 2018-07-27 | 2018-12-07 | 云南农业大学 | The Ravenna grass genoid EfNAC44 that Ravenna grass wild species are expressed by low temperature stress |
| CN108949780B (en) * | 2018-07-27 | 2019-07-30 | 云南农业大学 | The Ravenna grass genoid EfNAC44 that Ravenna grass wild species are expressed by low temperature stress |
| CN109197570A (en) * | 2018-10-11 | 2019-01-15 | 云南农业大学 | A method of improving sugarcane wild species resource pollen collection amount |
| CN109197570B (en) * | 2018-10-11 | 2021-09-10 | 云南农业大学 | Method for improving collection amount of sugarcane wild species resource pollen |
| CN110622811A (en) * | 2019-10-23 | 2019-12-31 | 广西壮族自治区农业科学院 | Single-plant and combined drought-resisting screening method for sugarcane seedlings |
| CN110881405A (en) * | 2019-11-27 | 2020-03-17 | 云南省农业科学院甘蔗研究所 | Method for exploring and utilizing cutting hand density breeding potential of sugarcane wild species |
| CN114009299A (en) * | 2021-12-15 | 2022-02-08 | 云南英茂糖业(集团)有限公司 | Mountain scale integrated planting method for improving per unit yield of sugarcane |
| CN114009299B (en) * | 2021-12-15 | 2022-09-20 | 云南英茂糖业(集团)有限公司 | Mountain scale integrated planting method for improving per unit yield of sugarcane |
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