CN103828678A - Method for quickly identifying pear rotting-resistant disease - Google Patents
Method for quickly identifying pear rotting-resistant disease Download PDFInfo
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- CN103828678A CN103828678A CN201410079119.2A CN201410079119A CN103828678A CN 103828678 A CN103828678 A CN 103828678A CN 201410079119 A CN201410079119 A CN 201410079119A CN 103828678 A CN103828678 A CN 103828678A
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Abstract
The invention discloses a method for quickly identifying pear rotting-resistant disease. The method includes utilizing a rule that 'lenticel relative area' of one-year-old pear dormant branches and disease attack degree of an artificially-inoculated tree are obviously in position correlation to select the one-year-old pear dormant branches, and grading the rotting-resistant disease of the pear by testing the 'lenticel relative area' of epidermis of the branches. Compared with the prior art, the method has the advantages of quickness, simplicity, convenience and high efficiency.
Description
Technical field
What the present invention relates to is a kind of authentication method of pear tree disease, in particular the anti-rot rapid identification method of a kind of pear tree.
Background technology
Fungus of Pear Canker Disease all has generation in the each main producing region of China's pears, occurs heavier with Xinjiang, northwest, North China and other places.According to current investigation, Xinjiang Kuerle delicious pear diseased plant rate has reached 50%～80%, and the crisp pears diseased plant rate in the Northwest reaches 30%～50%; North China's pear, Chinese pear diseased plant rate all reach more than 30%.Rot has that generation area is wide, the incidence of disease is high, unmanageable feature, the serious theatre of falling ill, and the sick scar of tree body is innumerable, limb is incomplete, even causes a large amount of Si Shuhuohui garden, and serious threat is to the sound development of China's pears industry.
Exploitation by new technology and multiple technologies integrated, sets up continuable pears rot Comprehensive Preventing control techniques system for planted in different ecological areas, promotes China's pears rot prevention and control technical merit although can be.But, cultivate and filter out the improved Varieties of anti-rot and carry out spread, be the only way which must be passed of effectively controlling Fungus of Pear Canker Disease and spreading the safety in production that increases the weight of, ensures China's pears.At present, the resistance of the different quality resources of qualification pear tree to rot, only gets annual resting shoot sample, adopts electric iron to scald Vitro Inoculation Technique, or field nursery stock is carved and hinders in-situ inoculating.Like this, not only need a large amount of bacterium source of activation culture, after inoculation, still need and wait for about one week, could statistical results.As can be seen here, for the anti-rot characterization and evaluation of pear tree, still lack quick, easy, effective method.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, provide a kind of pear tree anti-rot rapid identification method, to provide a kind of pear tree quick, easy, that effective method identifies high resistance type anti-rot kind.
The present invention is achieved by the following technical solutions, the present invention includes following steps:
The anti-rot rapid identification method of a kind of pear tree, comprises the following steps:
(1) get the annual resting shoot of pear tree, peel the epidermis of branch, measure supracutaneous hole skin and account for the area ratio of whole epidermis, obtain " hole skin relative area ";
(2) disease resistance classification: the Pear Trees with Various Cultivars of " hole skin relative area "≤2% is high resistance type, " hole skin relative area " Pear Trees with Various Cultivars of (2%, 6%) is intermediate form in open interval, the Pear Trees with Various Cultivars of " hole skin relative area " >=6% is susceptible type.
Preferably, in described step (1), annual resting shoot is the annotinous branch to the tree crown periphery before rudiment after pear tree fallen leaves.
Preferably, in described step (1), the middle part that the sampling point of annual resting shoot is branch.
Preferably, in described step (1), the area that strips of epidermis is 1～2 square centimeter.
Preferably, in described step (1), the assay method of " hole skin relative area " is: first with the above precision scanning of 300dpi epidermis, preserve imaging, measure total pixel value of epidermis and total pixel value of hole skin, the ratio of total pixel value of hole skin and total pixel value of epidermis is " hole skin relative area ".
Operation principle of the present invention is: the present invention has chosen various types of kind and represented 38 parts of resources (deriving from 12 kinds of pear), by traditional electrical flatiron scald, the resistance of Vitro Inoculation Technique method qualification different cultivars resource to rot, carry out correlation analysis by the measurement result of the aspect indexs such as morphology, cytology, physiology and molecular biology again,, between " the hole skin relative area " of the annual resting shoot of pear tree and the occurring degree of artificial infection pear tree, there is significant positive correlation (its correlation coefficient r=0.9381) in final discovery.Therefore, established the feasibility that substitutes the resistance level of traditional inoculation method qualification pear tree different cultivars resource to rot with " hole skin relative area ".
The present invention has the following advantages compared to existing technology: the invention provides the anti-rot rapid identification method of a kind of pear tree, the method only need strip the epidermis of the annual resting shoot of pear tree, measure " hole skin relative area ", can, by anti-pear tree rot qualitative classification, there is quick, easy, efficient feature; And traditional electric iron is scalded, Vitro Inoculation Technique method, before inoculation, need to activate a large amount of bacterial classifications, the branch quantity that inoculation needs is many, needs to cultivate and within 7～10 days, just can observe qualification after inoculation, and qualification cycle is long.
Embodiment
Below embodiments of the invention are elaborated, the present embodiment is implemented under taking technical solution of the present invention as prerequisite, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Embodiment 1
The present embodiment is chosen fresh pears in June, Nan Guoli and Pucheng three kinds of Pear Trees with Various Cultivars of snow and is represented resource, carries out anti-rot Rapid identification, comprises the following steps:
(1) get respectively after fallen leaves to the peripheral annual resting shoot of tree crown of the above-mentioned two kinds of Pear Trees with Various Cultivars before rudiment, peel the epidermis at branch middle part, described epidermis strip approximately 1 square centimeter of area, epidermis is fixed on blank sheet of paper with transparent viscose glue, repeat 5 times;
A, be fixed on the epidermis on blank sheet of paper with the scanner scanning of 300bpi precision, preserve imaging;
B, utilization photoshop image processing software, choose with square selection tool the unit epidermis area that hole skin is evenly distributed, and checks in the pixel value Sa of this unit epidermis area in " histogram " column in " image " menu;
" brightness " and " contrast " of c, increase image, select " magic wand menu ", adjust " tolerance " value, make " magic wand menu " just in time can select all hole skins and be advisable, then in " histogram " column in " image " menu, check in the pixel value La of this unit epidermis area endothelium hole area;
" hole skin relative area " RA(% of d, calculating branch epidermis), RA=La/Sa × 100%;
(2) disease resistance classification:
June fresh pears: La=806, Sa=43818, RA=1.84%, is high resistance type;
Nan Guoli: La=2436, Sa=43820, RA=5.56%, is intermediate form;
Pucheng snow: La=4710, Sa=44019, RA=10.70%, is susceptible type.
Embodiment 2
The present embodiment is chosen 38 parts of Pear Trees with Various Cultivars and is represented resource, relates to 12 kinds and a species hybrid in country variant and area, identifies the anti-rot degree of these 38 parts of Pear Trees with Various Cultivars resources, comprises the following steps:
(1) choose above-mentioned 38 parts of Pear Trees with Various Cultivars December and represent resource, get 3 of the peripheral annual resting shoots of tree crown, peel branch middle part epidermis epidermis strip approximately 2 square centimeters of areas, epidermis is fixed on blank sheet of paper with transparent viscose glue;
A, be fixed on the epidermis on blank sheet of paper with the scanner scanning of 300bpi precision, preserve imaging;
B, utilization photoshop image processing software, choose with square selection tool the unit epidermis area that hole skin is evenly distributed, and checks in the pixel value Sa of this unit epidermis area in " histogram " column in " image " menu;
" brightness " and " contrast " of c, increase image, select " magic wand menu ", adjust " tolerance " value, make " magic wand menu " just in time can select all hole skins and be advisable, then in " histogram " column in " image " menu, check in the pixel value La of this unit epidermis area endothelium hole area;
" hole skin relative area " RA(% of d, calculating branch epidermis), RA=La/Sa × 100%;
(2) disease resistance classification: the Pear Trees with Various Cultivars of " hole skin relative area "≤2% is high resistance type, " hole skin relative area " is in open interval (2%, 6%) Pear Trees with Various Cultivars is intermediate form, and the Pear Trees with Various Cultivars of " hole skin relative area " >=6% is susceptible type, and qualification result is as shown in table 1.
Meanwhile, the peripheral annual resting shoot of tree crown that represents resource taking above-mentioned 38 parts of Pear Trees with Various Cultivars, as test material, adopts electric iron scald, Vitro Inoculation Technique method as a control group.Every part of resource is got 5 of long 30～40 centimetres of branch sections, on the branch skin of every branch section, poker the wound of 3 diameter 5～7mm with electric iron, wound depth reaches xylem, the rotten pathogenic bacteria cake that wherein potato dextrose agar PDA cultivation 4 days for the inoculation of the wound of 4 branches, punching diameter are 5mm, another root branch wound is inoculated in contrast with aseptic PDA.Each branch section of processing is put in plastic preserving box, and aseptic moistening gauze moisturizing, was placed in 25 DEG C of constant temperature culture after 3 days, observed moisturizing and incidence every 24 hours, after inoculation, 10d measures expansion scab diameter, calculates each kind scab " spreading coefficient " Ec, represents each kind inoculation occurring degree:
Ec=∑ (expansion scab diameter-electric iron punching diameter)/morbidity hole count.
According to vaccination scab spread scenarios, these 38 parts of Pear Trees with Various Cultivars are represented to resource carries out disease resistance classification, be divided into altogether Three Estate:
High resistance type: Ec≤6;
Intermediate form: 6<Ec<50;
Susceptible type: Ec >=50
The present invention and traditional inoculated identification result are as shown in table 1 below:
Table 1: Pear Trees with Various Cultivars represents the anti-rot qualification result of resource
In table 1, can find out, the qualification result of the method for the invention and traditional electrical flatiron are scalded, the qualification result of Vitro Inoculation Technique method is in full accord, and recall rate reaches 100%; On the other hand, 7～10 days consuming time of traditional authentication method, only has about 1 hour and the method for the invention is consuming time, and therefore, than conventional method, the method for the invention advantage is remarkable.
Claims (5)
1. the anti-rot rapid identification method of pear tree, is characterized in that, comprises the following steps:
(1) get the annual resting shoot of pear tree, peel the epidermis of branch, measure supracutaneous hole skin and account for the area ratio of whole epidermis, obtain " hole skin relative area ";
(2) disease resistance classification: the Pear Trees with Various Cultivars of " hole skin relative area "≤2% is high resistance type, " hole skin relative area " Pear Trees with Various Cultivars of (2%, 6%) is intermediate form in open interval, the Pear Trees with Various Cultivars of " hole skin relative area " >=6% is susceptible type.
2. the anti-rot rapid identification method of a kind of pear tree as claimed in claim 1, is characterized in that, in described step (1), annual resting shoot is the annotinous branch to the tree crown periphery before rudiment after pear tree fallen leaves.
3. the anti-rot rapid identification method of a kind of pear tree as described in as arbitrary in claim 1 or 2, is characterized in that, in described step (1), and the middle part that the sampling point of annual resting shoot is branch.
4. the anti-rot rapid identification method of a kind of pear tree as claimed in claim 1, is characterized in that, in described step (1), the area that strips of epidermis is 1～2 square centimeter.
5. the anti-rot rapid identification method of a kind of pear tree as claimed in claim 1, it is characterized in that, in described step (1), the assay method of " hole skin relative area " is: first with the above precision scanning of 300dpi epidermis, preserve imaging, measure total pixel value of epidermis and total pixel value of hole skin, the ratio of total pixel value of hole skin and total pixel value of epidermis is " hole skin relative area ".
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107517726A (en) * | 2017-10-14 | 2017-12-29 | 中国科学院新疆生态与地理研究所 | A kind of fast quick access bacterium of canker of apple fruit pathogen and the method for pathogenicity Rapid identification |
| CN109001368A (en) * | 2018-06-11 | 2018-12-14 | 塔里木大学 | A method of detection Kuerle delicious pear tree infects rot disease |
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| CN102640671A (en) * | 2012-05-02 | 2012-08-22 | 湖北省农业科学院果树茶叶研究所 | Method for identifying Alternaria altemata (Fr.) Keissler resistance of pear germplasm |
| CN102719559A (en) * | 2012-06-12 | 2012-10-10 | 中国检验检疫科学研究院 | Chip for screening avsunviroid viroid and application of chip |
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Patent Citations (4)
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| CN103371062A (en) * | 2012-04-17 | 2013-10-30 | 河北省农林科学院昌黎果树研究所 | In-vitro identification method for venturia nashicola resistance |
| CN102640671A (en) * | 2012-05-02 | 2012-08-22 | 湖北省农业科学院果树茶叶研究所 | Method for identifying Alternaria altemata (Fr.) Keissler resistance of pear germplasm |
| CN102719559A (en) * | 2012-06-12 | 2012-10-10 | 中国检验检疫科学研究院 | Chip for screening avsunviroid viroid and application of chip |
| CN103503737A (en) * | 2013-10-09 | 2014-01-15 | 中国农业科学院郑州果树研究所 | Method for identifying resistance of cytosporacarphosperma |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107517726A (en) * | 2017-10-14 | 2017-12-29 | 中国科学院新疆生态与地理研究所 | A kind of fast quick access bacterium of canker of apple fruit pathogen and the method for pathogenicity Rapid identification |
| CN109001368A (en) * | 2018-06-11 | 2018-12-14 | 塔里木大学 | A method of detection Kuerle delicious pear tree infects rot disease |
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