CN101832943B - Quality detection method for pollen for pollinating Korla fragrant pears - Google Patents
Quality detection method for pollen for pollinating Korla fragrant pears Download PDFInfo
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Abstract
The invention discloses a quality detection method for pollen for pollinating Korla fragrant pears. Three-level detection is carried out on the pollen purity, the germination percentage and the fruiting rate of the Korla fragrant pears, and the method comprises the following steps of: physically judging whether the purity of the pollen to be detected is 80 to 100 percent and the impurity particles account for 0 to 15 percent under a microscope according to the principle of metrology; detecting the viability of the pollen population through a germination test, seeding the pollen in a culture medium, keeping moisture at 25 DEG C for four hours, and metering the germination rate of the pollen under the microscope, wherein the average germination rate of qualified pollen is more than or equal to 50 percent; and finally judging the inherent quality of the pollen by combining the emasculation with the artificial pollination technology and considering the inflorescence fruiting rate, the flower fruiting rate and the fruit texture feature of special pollen. The invention creatively combines laboratory analysis and field investigation and analysis, establishes a set of complete and strong-applicability quality detection method for the pollen for pollinating the Korla fragrant pears, and has high application value.
Description
Invention field
The present invention relates to pollen detection technique field.Specifically, the present invention relates to a kind of pollination and use the pear pollen technical field of quality detection.
Background technology
Pears are a kind of important temperate zone deciduous fruit trees, and great majority cultivation pears are wide in variety to be the cross-pollination type, must could guarantee its yield and quality by the suitable pollinizers of configuration during cultivation.The way that addresses this problem traditionally is interlacing or disposed adjacent pollinizers in same orchard, or on main breed the high position grafting pollinated variety.The development of agriculture industrialization has in recent years changed traditional pears planting patterns, and in order to improve planting benefit, many commercial cultivations theatre does not dispose pollinizers when building the garden, and the pollination problem solves through artificial pollination with pollen through buy special-purpose pollination at the florescence.
Pollination is a kind of important means of production with pear pollen, and as a kind of special living body biological resource, the detection technique difficulty of its quality is bigger.And the detection of pollen quality is significant; The one, pollination is circulated in reality on the agricultural materials market as a kind of commodity with pollen; The science detection of its quality can be the right dispute that causes because of quality problems provides legal services; The 2nd, in fact pollination is directly connected to the quality and yield of the operatic circle with the pollen quality, has influence on the sustainable development of pears industry, so a kind of research and development of pollinating with the science detection architecture of pear pollen are the desirabilities of current pears industrialized development.
Yet for a long time, the simple traditionally detection mode to the pollen germination rate is continued to use in the detection of pear pollen quality always.This kind mode can only be judged the viability of pollen, and to the seeds under the pollen and kind, being vacant to the back of pollinating with the affinity of main breed, the detection of pollen texture effect, and the evaluation result of making can not satisfy produces and the requirement in market.Therefore, need badly set up one the cover science pollination with pollen quality testing system.
Summary of the invention
The objective of the invention is to overcome the deficiency of existing quality detection method for pollen; A kind of science, the easy Kuerle delicious pear special-purpose pear pollen quality detection technology system of pollinating is provided; A kind of detection technique system of pollen quality can be provided for agriculture administrative service division, the inspection technology and the standard of qualified pollen also can be provided for relevant pollen manufacturing enterprise.
The invention provides a kind of conclusive evidence and quantivative approach of pollen for pollinating Korla fragrant pears quality testing.
Technical scheme of the present invention: pollen production that provides through the pollen manufacturer and quality inspection file are tentatively assert seeds and the kind name that supplies under the inspection pollen, and the quality testing when this kind pollen pollinate in order to Kuerle delicious pear is made up of laboratory detection and field detection.Main purity, form from agrobiology angular detection pollen detected in the laboratory, and the viability that detects pollen colony; It mainly is to combine the artificial pollination technology to realize through connecting to be castrated that the field is detected, and comes the final inherent quality of judging pollen through the fruit texture properties of considering inflorescence fruit-setting rate, flower fruit-setting rate and extraordinary pollen.
Particularly, the invention provides a kind of conclusive evidence and quantivative approach of pollen for pollinating Korla fragrant pears quality testing, step is following:
1, judge the purity that supplies inspection pollen through the metrology principle from form at microscopically, the index of main foundation is the form of pollination with conventional pears kind pollen.
2, pass through the vigor that the germination experiment detects pollen colony, the index of main foundation is that pollen is sowed on the nutrient culture media of special formulation, preserves moisture through specific double dish, under 25 ℃, cultivates 4h, at the germination rate of microscopically metering pollen.
3, combine the artificial pollination technology to realize through connecting to be castrated.Connect and by the theoretical foundation of emasculation technique to be: Kuerle delicious pear is strict entomophilous flower vegetation type, and the experiment proof has been got rid of the natural hybrization that causes because of hearsay and entomophila behind artificial removal's perianth, has exempted and has spent the loaded down with trivial details of bagging after preceding bagging, the pollination traditionally.Combine artificial pollination at last, come the final inherent quality of judging pollen through considering inflorescence fruit-setting rate, flower fruit-setting rate and fruit texture properties.
Further, it is following to the invention provides a kind of conclusive evidence and quantivative approach step of pollen for pollinating Korla fragrant pears quality testing:
1, will supply inspection pollen evenly to scatter on the microslide of thickness at 0.8-1.0mm, its mode of appearance is observed in (* 100) under the optical microscope.The standard of qualified pollen is: form is full, and pollen wall does not have and caves in ellipse or oval, three obvious germinal furrows of tool.Pollen pole axis 40 μ m ± 5 μ m, red axle 20 μ m ± 5 μ m.Pollen purity is at 80%-100%, and impurity particle is counted 0-15%.
2, the local pollen of newly adopting then can directly be used for pollen germination power and detects, and is the pollen that ships of pollen or outer ground warp cold chain of low tempertaure storage last year as supplying inspection pollen, and the processing that must under 25-30 ℃ of normal temperature, get damp again can be used for the detection of pollen germination power.The way of moisture regain is in an exsiccator, to put one glass of water, and pollen is dried in the air out on a parchamyn paper, places on the exsiccator pallet, then loam cake sealing 8h.Pollen germination detects the culture medium prescription that uses and is sucrose 10%, agar 1%, boric acid 0.03%, and the preparation water is a distilled water.Must heat and stir 105 ℃ of sterilizations of high temperature 0.5h in the nutrient culture media process for preparation with glass rod.Select bottom wall thickness＜1mm double dish for use, culture medium layer thickness＜2mm evenly is sowed at pollen in the nutrient culture media with hairline; Application rate is controlled at each visual field 100 ± 20 (* 100); Place under 25 ℃ of room temperatures and cultivate 4h,, add the double dish lid at the percent (* 100) of microscopically mensuration pollen germination.Whether constantly fine setting focusing during detection detects each pollen tube and extends under different focal.Measure 10 visuals field at random, computation of mean values, the average pollen germination rate of qualified pollen should be >=50%.
3, select the calm time of calm morning and evening in the bergamot pear balloon phase, pinch anthocaulus, perianth is extractd along calyx end ring shape with right hand thumb and forefinger nail with left hand; Only deposit ovary and column cap, i.e. gynoecium flower part, i.e. column cap, style, placenta and ovary; Column cap is awarded for inspection pollen with cotton swab or cigaratte filter head; Each No. of inflorescences pollination flower is counted 2-3, selects ray flower to castrate and pollinate, and middle disc flower is dredged removed.Every kind of pollen is established 10 repetitions, individual plant sub-district or single sub-district, each processing >=50 inflorescence altogether.Investigate each in early July (June physiological fallen fruit after) and handle field flower fruit-setting rate and inflorescence fruit-setting rate, ask its average.Pollination bergamot pear fruit is answered typical breediness of tool bergamot pear and commodity fruit characteristic: spindle, and fruit mean fruit weight >=120g, the natural calyx rate of taking off is more than 75%, and superfine is rate >=80% really.Inflorescence fruit-setting rate >=70% of qualified pollen, flower fruit-setting rate >=30%.
The computing formula of the pollen germination rate that adopts among the present invention: pollen frequency * 100% in pollen germination rate=each pollen tube growth pollen number/visual field, the visual field.
The computing formula of the inflorescence fruit-setting rate that adopts among the present invention: the inflorescence fruit-setting rate=No. of inflorescences of bearing fruit/pollination inflorescence sum * 100%.
The computing formula of the flower fruit-setting rate that adopts among the present invention: the sum of bearing fruit of flower fruit-setting rate=each processing/(each handles pollination flower sum) * 100%.
The natural calyx rate phase computing formula of taking off that adopts among the present invention: natural calyx rate=take off calyx fruit number/100 fruit picking at random * 100% that takes off.
The computing formula of the pollen purity that adopts among the present invention is: total number of particles * 100% in pollen purity=qualified pollen number/visual field; Impurity particle is counted total number of particles * 100% in percent=non-POLLEN MORPHOLOGY total number of particles/visual field
Adopt the superfine fruit of bergamot pear operative norm to be among the present invention: NY/T585-2002, prominent top fruit is no more than 10%, and single fruit weight is at 120-160g, no scurf, malformed fruit, the fruit face cleans, and carpopodium is complete.
Computing method, formula that above the present invention adopts, and the standard of carrying out all be that this area is used always, also is scope well known to those of ordinary skill in the art.
The above-mentioned artificial pollination method that the present invention relates to is that Dan Huadian awards mode.Pollinate and also can as tremble the mode of awarding through special-purpose pollination device of knapsack sprayer, pears or manual work.The proportioning of pollen and filler is undertaken by routine with the pollination operational method.
Through the concrete summary of the invention of embodiment of the present invention, can reach following effect:
The present invention through from examined three aspects of pollen purity, vigor and kind (pollen texture) realized to pollination with before the pollen pollination with pollination after whole process and the complete detection of mode of appearance and inherent kind property; Test item is thorough; Trace routine is complete; Testing process had both taken into full account the singularity of Kuerle delicious pear to the requirement of pollination pollen; Also taken into account the pollination of present China with the pollen preparation and manage real national conditions--partial pollen is that florescence last year is prepared, preserved a growth season at low temperatures.Simultaneously; The invention property ground identifies POLLEN MORPHOLOGY and the lab analysis of the ARTIFICIAL CULTURE of pollen and the field investigation analysis combination of fruit-setting rate investigation and features of fruits investigation, set up one overlap the strong pollen for pollinating Korla fragrant pears quality testing of complete applicability conclusive evidence and quantivative approach.
2. detection technique is easy, suitable; The general agrotechnical centralab at county level of experimental facilities required for the present invention and material possesses fully; Detection method is quick, convenient, and in the present invention, first and second step is detected to pollinate and accomplished in preceding 1-2 days; The 3rd step can accomplish in mid-July, was suitable for the real-time monitoring to the pollen product quality of market control departments, agriculture administrative service division and pollen manufacturing enterprise.
3. do not carried out under the prerequisite of artificial bagging with connecting by the castration method among the present invention; The fruit-setting rate that detects extraordinary pollen through artificial pollination is a science; Also be reliably, this method has been removed the loaded down with trivial details of artificial bagging traditionally from, is an innovation; Not only can detect in order to pollen, this technology also can be widely used in the crossbreeding work to increase work efficiency.
Description of drawings
Fig. 1 is the sowing synoptic diagram of pollen on nutrient culture media when supplying the inspection pollen viability to detect, among the figure, 1 be pollen granule, 2 for double dish lid, 3 be that nutrient culture media, 4 is a double dish.
Fig. 2 is a balloon phase pears list flower morphology of tea flower organ synoptic diagram, and among the figure, 5 is that perianth, 6 is that column cap, 7 is ovary.
Fig. 3 carries out manual work to balloon phase pears list flower to connect by castration back female flower device form synoptic diagram, and among the figure, 8 is that style, 9 is placenta.
Embodiment
Below, lift embodiment the present invention is described, still, the present invention is not limited to following embodiment.In addition, in following explanation, if no special instructions, % all refers to the m/m mass percent.
The major equipment and the material that relate among the present invention have:
Equipment: ordinary optical microscope, microslide, counter, double dish, exsiccator, high-pressure sterilizing pot, illumination box
Material: sucrose (chemical pure), agar (chemical pure), boric acid (chemical pure), distilled water
All reagent selected for use among the present invention and instrument all are well known selecting for use, but do not limit enforcement of the present invention, and other reagent more well known in the art and equipment are all applicable to the enforcement of the following embodiment of the present invention.
Embodiment one:
1, will supply inspection pollen evenly to scatter on the microslide of thickness at 0.8-1.0mm, its form is observed in (* 100) under optical microscope.Select the suitable visual field, adjust focal length, a particle in the visual field is carried out form judge.Qualified POLLEN MORPHOLOGY is full, and pollen wall does not have and caves in, ellipse or oval, and three obvious germinal furrows of tool, pole axis 40 ± 5 μ m, red axle is at 20 ± 5 μ m.Qualified pollen purity should be more than 80%, and the impurity particle number is lower than 15%.Supply the every kind pollen field of view of inspection to be not less than 10.The computing formula of pollen purity is: total number of particles * 100% in pollen purity=qualified pollen number/visual field.Impurity particle is counted total number of particles * 100% in percent=non-POLLEN MORPHOLOGY total number of particles/visual field.
2, pollen germination detects the culture medium prescription use and is sucrose 10%, agar 1%, boric acid 0.03%, and the preparation water is a distilled water.Must heat and stir high temperature (105 ℃) sterilization 0.5h in the nutrient culture media process for preparation with glass rod.Select bottom wall thickness＜1mm double dish for use, culture medium layer thickness＜2mm evenly is sowed at pollen in the nutrient culture media with hairline, and application rate is controlled at each visual field 100 ± 20 (* 100), adds the double dish lid.Place illumination box to cultivate 4h down, measure pollen germination rate (* 100) at microscopically at 30 ℃.Whether constantly fine setting focusing during detection detects each pollen tube and extends under different focal.Measure 10 visuals field at random, computation of mean values.The average pollen germination rate of qualified pollen should be >=50%.
3, select calm time morning and evening in the bergamot pear balloon phase, pinch anthocaulus with left hand, right hand thumb and forefinger nail are extractd perianth along calyx end ring shape, only deposit the gynoecium flower part, i.e. column cap, style, placenta and ovary.Column cap is awarded for inspection pollen with cotton swab or cigaratte filter head, and each No. of inflorescences pollination flower number is not less than 3, selects ray flower to castrate and pollinate, and middle disc flower is dredged removed.With pollination device or tremble the mode of awarding and carry out artificial pollination and also can.Every kind of pollen is established 10 repetitions, individual plant sub-district or single sub-district, each processing >=50 inflorescence altogether.Each handles field flower fruit-setting rate and inflorescence fruit-setting rate in the early July investigation, asks its average.Pollination bergamot pear fruit is answered typical breediness of tool bergamot pear and commodity fruit characteristic: spindle, fruit mean fruit weight >=120g takes off the calyx rate more than 75%, one-level fruit rate >=80%.Inflorescence fruit-setting rate >=70% of qualified pollen, flower fruit-setting rate >=20%.
Embodiment two:
Well known, castrating and overlapping bag technique is two basic fundamentals commonly used in the artificial hybridization breeding.One of purpose of emasculation technique is the technology that prevents natural selfing when being artificial hybridization both sexes complete flower, and the purpose of cover bag technique then is to prevent the natural hybrization that wind pollination or insect pollination cause.The floral organ of pears is complete flowers, and most of kind shows as different flowering plant, and the pollination self fruit-setting rate is extremely low, like bergamot pear less than 2%.Generally to combine bagging to prevent to take place the natural hybrization that wind pollination or insect pollination cause to spending of castrating traditionally.The present invention is employed in the flower bud phase, i.e. the balloon phase, even castrated through manual work, because this fashion of pear flower is open, flower pesticide also ftractures, and has got rid of the possibility of natural hybrization and selfing fully.The present invention adopts the flower to castrating not carry out bagging; Under connecting by the castration condition; For the pears of insect pollination, the possibility that natural hybrization takes place is extremely low, reason be castrate insect pollinators such as back honeybee class and fly class can't be on achlamydeous placenta for want of enough anchorage forces and landing; Second as far as entomophila; Pollen density is in theory very low in the air, and the probability that drops on the column cap is lower, can't satisfy the required sprouting pollen number of pears normal development (wanting＞6 pollen/list flower at least) fully.
The present invention is in order to check this technological reliability; Confirm our above-mentioned supposition; Implemented following field experiment: the one, the single visit capacity of spending in the unit interval in back is castrated in investigation, and the 2nd, added up the natural setting percentage of inflorescence that does not carry out pear flower under the bagging prerequisite after castrating.Above-mentioned two experiments are not all to castrate as contrast.Relevant result is as shown in the table
The artificial company of table 1 bergamot pear and pear castrated back insect visit capacity statistics
The artificial company of table 2 bergamot pear and pear castrated back insect visit capacity statistics
Result through table 1 and 2 confirms, is not being carried out under the prerequisite of artificial bagging by the castration method with connecting among the present invention, and the fruit-setting rate that detects extraordinary pollen through artificial pollination is a science, also is reliable.This method has been removed the loaded down with trivial details of artificial bagging traditionally from, is an innovation, not only can detect in order to pollen, also can be widely used in the crossbreeding work to increase work efficiency.
Claims (1)
1. the method for a pollen for pollinating Korla fragrant pears quality testing through the bergamot pear fruit-setting rate after pollen purity, germination percentage and the artificial pollination is carried out three grades of detections, is characterized in that, it is following with pear pollen quality determining method step to pollinate:
(1) microscopically through the metrology principle from form judge to supply the purity of inspection pollen, to be detected drying supply inspection pollen with the writing brush uniform broadcasting on the microslide of thickness 0.8-1.0mm, place observation under the optical microscope; Supply inspection pollen should have the typical surface of pear pollen: form is full, and pollen wall does not have and caves in ellipse or oval; Three obvious germinal furrows of tool, pole axis 40 ± 5 μ m, red axle is at 20 ± 5 μ m; Pollen granule accounts for seized granule number ratio, and promptly pollen purity is at 80%-100%; Described pollen is the pollen that ships of pollen or outer ground warp cold chain of low tempertaure storage last year, should after moisture regain is handled under the 25-30 ℃ of normal temperature, be used for pollen germination power and detect; Described moisture regain is in an exsiccator, to put one glass of water, and pollen is dried in the air out on a parchamyn paper, places on the exsiccator pallet, then loam cake sealing 8h;
(2) through the vigor of the experiment detection pollen colony of germinateing, select nutrient culture media to pour double dish into, bottom wall thickness＜1m; Be paved into＜skim of 2mm thickness, cooling evenly is sowed at pollen in the nutrient culture media with hairline; Application rate is controlled at each visual field 100 ± 20 (* 100), adds a cover, and places under 25 ℃ of room temperatures and cultivates 4h; Measure pollen germination rate at microscopically, the average pollen germination rate of qualified pollen should be >=50%; Described culture medium prescription is sucrose 10%, agar 1%, boric acid 0.03%, and the preparation water is distilled water;
(3) combine artificial pollination method to realize through connecting to be castrated, select calm time morning and evening, pinch anthocaulus with left hand in the bergamot pear balloon phase; With right hand thumb and forefinger nail perianth is extractd along calyx end ring shape; Only deposit ovary and column cap, column cap is awarded for inspection pollen with cotton swab or cigaratte filter head, each No. of inflorescences pollination flower is counted 2-3; select ray flower to castrate and pollinate, middle disc flower is dredged removed; Every kind of pollen is established 10 repetitions, individual plant sub-district or single sub-district, each processing >=50 inflorescence altogether; Each handles field flower fruit-setting rate and inflorescence fruit-setting rate in the early August investigation; Ask its average, pollination bergamot pear fruit is answered typical breediness of tool bergamot pear and commodity fruit characteristic: spindle, fruit mean fruit weight >=120g; Take off the calyx rate more than 75%; One-level fruit rate >=80%, inflorescence fruit-setting rate >=70% of qualified pollen, flower fruit-setting rate >=30%.
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| CN103421737B (en) * | 2013-06-04 | 2015-04-15 | 塔里木大学 | Culture medium for promoting germination of Amygdalus L. pollen, culture method and application thereof |
| CN103983644A (en) * | 2014-03-10 | 2014-08-13 | 云南农业大学 | Determination method for tomato pollen vitality |
| CN106718833B (en) * | 2016-12-07 | 2019-06-07 | 延边朝鲜族自治州农业科学院（延边特产研究所） | Apple pear essence pollen pollinating method |
| CN111543308A (en) * | 2020-05-06 | 2020-08-18 | 中国农业科学院蜜蜂研究所 | Preparation method of pollen for pollination |
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| JP4177494B2 (en) * | 1997-10-24 | 2008-11-05 | 富士シリシア化学株式会社 | Pollen extender, artificial pollination method, and pollen preservation method |
| JP2001264234A (en) * | 2000-03-17 | 2001-09-26 | Matsushita Electric Ind Co Ltd | Pollen detection device |
| CN1994060A (en) * | 2006-01-02 | 2007-07-11 | 田宝江 | A fruit tree pollination method |
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| 何天明等.新疆香梨产区外源授粉花粉萌芽力检测.《新疆农业大学学报》.2010,第33卷(第2期),第120-123页. * |
| 何天明等.香梨花器特征及花粉生活力研究初报.《新疆农业大学学报》.2000,第23卷(第3期),35-38. * |
| 刘武林.花粉的采集、贮藏和生活力的测定.《植物学通报》.1985,第3卷(第3期),8-12. * |
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