阅读说明：本技术 一种甘蔗实生苗移栽前梢腐病接种胁迫的方法 (A kind of method that cane seedlings transplant preceding top rot inoculation stress ) 是由 黄海荣 杨荣仲 徐林 李翔 邓宇驰 经艳 王伦旺 唐仕云 周会 高轶静 谭芳 于 2019-09-30 设计创作，主要内容包括：本发明公开了一种甘蔗实生苗移栽前梢腐病接种胁迫的方法。该方法是在将甘蔗实生苗假植育苗穴盘中,待生长至6~8片叶时,将配制好的梢腐病孢子悬浊液喷淋甘蔗实生苗上部,覆盖塑料薄膜保温保湿,进行梢腐病接种,用于筛选出抗梢腐病单株或组合。该方法可以大规模接种实生苗,评价组合,提早淘汰感病单株,从而提高育种效率,减少误选几率,节省后期的劳力物力地力。本发明中的接种装置,可自动均匀进行甘蔗实生苗梢腐病接种,有效控制接种环境温度和湿度,提高了接种效果,减少了抗梢腐病甘蔗苗品种误选几率,可用于准确筛选抗梢腐病甘蔗品系。(A kind of method of top rot inoculation stress before being transplanted the invention discloses cane seedlings.This method is planted in seedling culture hole plate by cane seedlings, it is to be grown to 6 ~ 8 leaves when, prepared top rot spore suspension is sprayed into cane seedlings top, plastic covering film thermal insulating moisturizing, top rot inoculation is carried out, for filtering out anti-top rot single plant or combination.This method can be evaluated combination, be eliminated susceptible single plant ahead of time with large-scale inoculation seedling, to improve breeding efficiency, probability is falsely dropped in reduction, save labour's material resources soil fertility in later period.Classification inoculation apparatus in the present invention, can automatic uniform carry out the inoculation of cane seedlings top rot, effectively control inoculation environment temperature and humidity, improve effect of inoculation, reduce anti-top rot sugarcane seedling kind and falsely drop probability, can be used for accurately screening anti-top rot sugarcane line.)

1. a kind of method of top rot inoculation stress before cane seedlings are transplanted, which comprises the steps of:

(1) cane seedlings are cultivated；

(2) it after cane seedlings germination and growth to 2 ~ 3 true leaves, is planted in seedling culture hole plate；

(3) prepared by top rot spore suspension: acquisition sugarcane top rot leaf collects top rot pathogen spore, is put into PDA culture Then base culture uses sterile water and polysorbas20 to configure spore concentration as 10⁵~10⁶The top rot spore suspension of a spore/ml, The mass percent of tween is 0.05% ~ 1.5% in the sterile water and tween；

(4) it is inoculated with: when the cane seedlings wait be planted grow to 6 ~ 8 leaves, prepared top rot spore suspension being sprayed Cane seedlings top, plastic covering film thermal insulating moisturizing 24 hours.

The method of top rot inoculation stress before 2. cane seedlings according to claim 1 are transplanted, which is characterized in that described The mass percent of polysorbas20 is 0.1% in sterile water and polysorbas20.

The method of top rot inoculation stress before 3. cane seedlings according to claim 1 are transplanted, which is characterized in that described Step (4) is carried out in cane seedlings top rot classification inoculation apparatus, the cane seedlings top rot classification inoculation apparatus, including pedestal, Top cover, reticular lamina and support rod, the reticular lamina be it is horizontally disposed, it is netted that the support rod connects pedestal, multilayer from bottom to top Plate and top cover constitute inoculation frame, and the top cover is surrounded by spool, and the spool periphery is wound with window-blind, the inoculation frame one Side is equipped with inoculation liquid storage box, and the inoculation liquid storage box connects water outlet pump input terminal by connecting tube, and the water outlet pump is defeated The connection of one end of outlet and outlet pipe, the outlet pipe extend along inoculation frame, and the other end is connect with atomizer, multiple mists Change the top that spray head is respectively arranged on each reticular lamina.

The method of top rot inoculation stress before 4. cane seedlings according to claim 3 are transplanted, which is characterized in that described It is equipped with heater and humidifier in pedestal, is equipped with supply air system in the heater and humidifier.

The method of top rot inoculation stress before 5. cane seedlings according to claim 4 are transplanted, which is characterized in that described Step (4) inoculation specific steps include:

(1) hole tray for being planted cane seedlings is placed on the reticular lamina in inoculation frame, puts down window-blind；

(2) prepared top rot spore suspension is poured into inoculation liquid storage box, starts water outlet pump, top rot spore is hanged Turbid pumps to each atomizer, is equably sprayed on cane seedlings；

(3) start heater and humidifier, set temperature is 25 ~ 35 DEG C and relative humidity 70 ~ 90%, creates spore germination and invades Contaminate the favorable environment of blade.

The method of top rot inoculation stress before 6. cane seedlings according to claim 5 are transplanted, which is characterized in that described Top rot spore turbid liquid spraying flow is 3 ~ 5ml/min, sprays 1 ~ 2min, can tie drop with sugarcane lobus cardiacus folic pocket and be preferred.

7. application of the method as claimed in any one of claims 1 to 6 in the anti-slightly maize ear rot breed breeding of cane seedlings.

8. application according to claim 7, it is characterised in that: inoculation starts to observe and record sugarcane ending crease blade after 5 days Contract the lesion situation distorted, adds up disease plant number after 30 days, determines the susceptible situation of cane seedlings, and then preliminary screening is anti-out The sugar cane breed of top rot.

Technical field

The invention belongs to the anti-slightly maize ear rot breed breeding fields of sugarcane, and in particular to a kind of preceding top rot of cane seedlings transplanting The method for being inoculated with stress.

Background technique

Sugarcane top rot is a kind of gas borne fungus diseases, and the conidium of the germ is brought by air-flow to sugarcane toppers head lobe piece On, sugarcane is infected in rudiment under preference temperature and humidity.It is yellow that the Sugarcane Leaves of infection top rot have mainly shown blade chlorisis Change, shrinkage distortion, eventually lead to plant stop growing or on taper tender tissue formed top rot shape.The high temperature of China 7-9 month High humidity weather is the main generation period of top rot, sufficiently pours water after arid, encounters rainfall after applied nitrogen is excessive or arid, all The extensive generation of top rot can be caused, 1989, the outburst of Pearl River Delta sugarcane district POJ2878 top rot caused 10%~38% sugarcane stem is withered.The test of 2012~2014 years Guangxi sugar cane breed region and 2013~2014 years national sugarcanes Regional experiment of variety investigation, kind disease incidence reach 100%.Disease can lead to the sugarcane underproduction 5%~20%, cane sugar It reduces up to 3%.Top rot has the characteristics that onset speed is fast, propagated strong, such as controls the sprawling of disease not in time, will be serious Influence the sustainable development of Sugarcane Industry.

The prevention and treatment most economical effective method of top rot is to the progress top rot inoculation of sugarcane seedling and to carry out disease-resistant variety breeding, The seedling (vegetative propagation) that the inoculation technique of sugarcane slightly maize ear rot cultivates growth generally directed to sugarcane stem eye is inoculated with, the side of inoculation Method is to inject sick liquid in seedling base portion leaf sheath folic pocket injection or needle point method, this inoculation technique is claimed not to be suitable for extensive sugarcane seedling Inoculation.Current sugarcane slightly maize ear rot is inoculated with two methods: 1) seedling (vegetative propagation) of growth is cultivated for sugarcane stem eye It carries out, the method can only carry out a small amount of sugar cane breed to evaluate preferably disease-resistant；2) the small seed for cultivating growth to stem eye is arrived Field, growth nature is susceptible, to eliminate susceptible variety.But the method may be in some plot of some time because of weather or ring This is not susceptible for the factor in border, qualitative disease-resistant variety and falsely drop the breeding and growth into second year, cause manpower soil fertility financial resources Waste, breeding efficiency are low.

Summary of the invention

A kind of method of top rot inoculation stress before being transplanted the purpose of the present invention is to provide cane seedlings, in sugarcane reality Raw seedling-breeding early stage, is inoculated with, and carry out Resistance Identification using spray slightly maize ear rot spore suspension, can be grown directly from seeds with large-scale inoculation Seedling, evaluation combination, eliminates susceptible single plant ahead of time, to improve breeding efficiency, probability is falsely dropped in reduction, saves labour's material resources in later period Soil fertility.

To achieve the above object, a kind of method of top rot inoculation stress before being transplanted the present invention provides cane seedlings, Include the following steps:

(1) cane seedlings are cultivated: sugarcane hybrid seed is seeded in sugarcane seed plate, under greenhouse, and control temperature 28~35 DEG C of degree, relative humidity 85~90%, seed germination, growth；

(2) it sugarcane heeled-in seedling: after cane seedlings germination and growth to 2~3 true leaves, is planted in seedling culture hole plate；

(3) prepared by top rot spore suspension: acquisition sugarcane top rot leaf collects top rot pathogen spore, is put into PDA Then culture medium culture uses sterile water and polysorbas20 to configure spore concentration as 10⁵~10⁶The top rot spore of a spore/ml is outstanding Turbid, the mass percent of tween is 0.05%~1.5% in the sterile water and tween；

(4) it is inoculated with: when the cane seedlings wait be planted grow to 6~8 leaves, by prepared top rot spore suspension Cane seedlings top is sprayed, Sugarcane Leaves is sufficiently humidified so as to substantially and contains disease liquid, disease liquid is made to be deposited in young leaves tender shoots position； Plastic covering film thermal insulating moisturizing 24 hours, creates an enabling environment as spore germination and infects blade.

Preferably, in above-mentioned technical proposal, the mass percent of polysorbas20 is 0.1% in the sterile water and polysorbas20, The ability that tween makes bacterium solution be adhered blade base is stronger, improves effect of inoculation.

Preferably, in above-mentioned technical proposal, the step (4) carries out in cane seedlings top rot classification inoculation apparatus, described Cane seedlings top rot classification inoculation apparatus, the cane seedlings top rot classification inoculation apparatus, including pedestal, top cover, reticular lamina with Support rod, the reticular lamina be it is horizontally disposed, the support rod connect from bottom to top pedestal, multilayer reticular lamina and top cover composition connect Kind frame, the top cover are surrounded by spool, and the spool periphery is wound with window-blind, and the inoculation frame one side is equipped with inoculation liquid Storage box, the inoculation liquid storage box connect water outlet pump input terminal, the water outlet pump output terminal and outlet pipe by connecting tube One end connection, the outlet pipe extend along inoculation frame, and the other end is connect with atomizer, and multiple atomizers are respectively arranged on The top of each reticular lamina.

Preferably, in above-mentioned technical proposal, heater is equipped in the pedestal of the cane seedlings top rot classification inoculation apparatus And humidifier, the heater and humidifier are interior equipped with supply air system.

Preferably, in above-mentioned technical proposal, what the step (4) was inoculated in cane seedlings top rot classification inoculation apparatus Specific steps include:

(1) hole tray for being planted cane seedlings is placed on the reticular lamina in inoculation frame, puts down window-blind；

(2) prepared top rot spore suspension is put into inoculation liquid storage box, starts water outlet pump, by top rot spore Sub- suspension pumps to each atomizer, is equably sprayed on cane seedlings；

(3) start heater and humidifier, set temperature is 25~35 DEG C and relative humidity 70~90%, creates spore and sprouts Send out and infect the favorable environment of blade.

Preferably, in above-mentioned technical proposal, the top rot spore turbid liquid spraying flow be 3~5ml/min, spray 1~ 2min can tie drop with sugarcane lobus cardiacus folic pocket and be preferred.

The method of top rot inoculation stress is in the anti-slightly maize ear rot breed breeding of cane seedlings before above-mentioned cane seedlings are transplanted In application.

Preferably, in above-mentioned application, inoculation started to observe and record the lesion situation of sugarcane ending leaf-shrinkage distortion after 5 days, Add up disease plant number after 30 days, determines the susceptible situation of cane seedlings, and then preliminary screening goes out the sugar cane breed of anti-top rot.

Compared with prior art, the invention has the following beneficial effects:

1. the present invention can carry out extensive seedling inoculation slightly maize ear rot using spray inoculation, inoculation efficiency is higher, more succinctly, Cane breeding mechanism can be served, resists slightly maize ear rot breeding for cane seedlings.

2. the present invention is inoculated with cane seedlings, susceptible single plant and combination can be eliminated ahead of time, reduce anti-top rot Sugarcane seedling kind falsely drops probability, improves breeding efficiency, reduces the investment of financial resource and material resource.

3. classification inoculation apparatus can automatic uniform progress cane seedlings top rot inoculation, effectively control inoculation environment in the present invention Temperature and humidity for top rot spore germination and infects blade and provides control environment, improves effect of inoculation, can be used for accurately sieving Select anti-top rot sugarcane line.

Detailed description of the invention

Fig. 1 is cane seedlings top rot classification inoculation apparatus structural schematic diagram of the invention.

Main appended drawing reference explanation:

1- pedestal, 2- top cover, 3- reticular lamina, 4- support rod, 5- spool, 6- window-blind, 7- inoculation liquid storage box, 8- water outlet Pump, 9- outlet pipe, 10- atomizer, 11- heater, 12- humidifier.

Specific embodiment

Specific embodiments of the present invention will be described in detail below, it is to be understood that protection scope of the present invention is not It is restricted by specific implementation.

Choose parent's Y63 (osmanthus sugar 94-119 × Guangdong sugar 93-159) sensitive to slightly maize ear rot, Y101 (Guangdong sugar 93-159 × Guangdong sugar 91-976), the anti-sense (disease-resistant and susceptible) (Guangdong sugar 94-128 × osmanthus sugar 00-122) of Y101, disease-resistant combination Y216 (YC97-40 × ROC22) and Y87 (Guangdong sugar 91-976 × ROC22) 5 combinations are used as subjects.