CN103749230A - Anti-rice black-streaked dwarf virus field inoculation identification method for rice varieties - Google Patents
Anti-rice black-streaked dwarf virus field inoculation identification method for rice varieties Download PDFInfo
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Abstract
The invention provides an anti-rice black-streaked dwarf virus field inoculation identification method for rice varieties, and belongs to the technical field of agricultural science. A piece of rice field with certain conditions is selected as an identification nursery, different rice varieties are sown in special stages, it is identified that virus transmission interbody small brown rice planthoppers in the nursery have certain virus carried rate and population density after wheat harvest, and after rice grows to a specific growth period, the anti-rice black-streaked dwarf virus level of the rice varieties is evaluated according to the severity degree of disease. With the method, anti-rice black-streaked dwarf virus identification can be performed on a large number of rice varieties at a time, and the defects that the manual identification scale in a laboratory and the identification variety number are small are overcome. The identification method can be applied to the multiple fields of screening of anti-rice black-streaked dwarf virus rice varieties, variety resistance evaluation, resistant variety breeding, researching on the law of genetics of resistance and the like.
Description
Technical field
The present invention relates to the field identification method of a kind of rice varieties to black streak dwarf resistance, belong to agricultural cience and farming techniques field.
Background technology
Black streaked dwarf virus of rice is a kind of virus disease being caused by rice black-streaked dwarf virus (Riceblack-streakeddwarfvirus, RBSDV), mainly by small brown rice planthopper, with persistence, without ovum mode, is propagated.In recent years, along with variation, the winter climate of cropping system and planting type warm and the spread of susceptible variety, pass virus mediator small brown rice planthopper generating capacity rising, black streaked dwarf virus of rice in Jiangsu, Zhejiang, Jiangxi and Fujian occurs on a large scale, and rise to rapidly and produce upper topmost virus disease.According to Jiangsu Province's plant protection unit, add up, end 2007, disease is coastal in this province, the Huaibei and in the ground such as Yancheng, Lianyun Harbour, Taizhou, Huaian, Yangzhou and the Xuzhou of going to river seriously occur, the area that harm occurs then reaches 30.8 ten thousand mu, the generation area of black streaked dwarf virus of rice in 2008 exceedes 4,000,000 mu, and disease outburst, popular situation are very severe.
Screening disease-resistant variety is applied to and produces is to prevent and treat the most directly effective method of black streaked dwarf virus of rice, also there is no up to now resistant variety application on producing.Field small brown rice planthopper often carries (the Ricestripevirus of stripe virus disease simultaneously, RSV) and RBSDV, but due to the extensive popularization of anti-stripe virus disease kind in recent years, make this disease obtain effective containment, therefore, the field test result of black streak dwarf can not be subject to the interference of RSV.In addition, the artificial authentication step in laboratory is comparatively loaded down with trivial details, identify that space is also restricted, can not carry out large-scale variety identification, and resource screening and resistive breeding often need to identify thousands of parts of rice varieties or strain, investigate the growing state of these kinds in field simultaneously, this makes the screening of anti-black streak dwarf paddy rice resource and breeding for disease resistance work be subject to great restriction, in the urgent need to inventing a kind of field identification method that can carry out on a large scale black streaked dwarf virus of rice evaluation, for the Resource exploitation screening of Rice Resistance black streak dwarf, breed breeding and genetic development research provide basic research means.
Summary of the invention:
Technical problem
The invention provides the Field inoculation authentication method of a kind of rice varieties to black streaked dwarf virus of rice resistance.
Technical scheme
The Field inoculation authentication method of rice varieties to black streaked dwarf virus of rice resistance, is characterized in that:
1) identify garden selection:
Select the continuous field of retransmitting above black streaked dwarf virus of rice for 3 years, and susceptible check variety of prior year is greater than 40% not preventing and treating the incidence of disease under condition, and after wheat harvest, identifies that in garden, passing virus mediator small brown rice planthopper is with malicious rate higher than 4%, insect density is up to 1.5 × 10
8head/hm
2, minimum is 6 × 10
6head/hm
2field as identifying garden, identify that garden surrounding plantation wheat entrusts one's child to the care of sb. district as small brown rice planthopper, to guarantee to obtain enough worms source;
2) broadcast cultivation mode
Participate in the variety seeds of evaluation, sowing in 15-20 days before wheat harvest, each material to be identified is broadcast 150 dry seedses, uniform broadcasting 1 row, the long 50cm of row, line-spacing 10cm; Sowing finishes rear field immersion 1-2 days, and weed killer herbicide is closed down, normal management after emerging, not with medicament insect protected between nursery period;
Length of time rice seedlings grow 5-6 transplants during leaf, and single seedling is planted, distance between rows and hills 15cm × 20cm, and each kind and family are planted 100 caves, conventional rich water quality management, any agricultural chemicals is not used in land for growing field crops, twice repetition;
3) small brown rice planthopper density
For small brown rice planthopper moves to the high-incidence season of identifying garden from wheatland, average, consistent in order to make to identify the small brown rice planthopper density of each point in garden before and after wheat harvest, every mornings 94 of point, afternoons are driven the small brown rice planthopper of identifying in garden with bamboo pole;
After identifying garden wheat harvest around, within 2-4 days, investigate small brown rice planthopper density: whole evaluation garden is adopted to 5 samplings of diagonal method, and 0.5m × 0.2m is looked in every some bat; Water moistening rectangle 35cm × 50cm white enamel dish inwall, aims at rice seedling middle and lower part, continuous shooting three times, and each bat looked into the rear white enamel dish that cleans of counting, carries out clapping next time and looks into;
4) small brown rice planthopper is with malicious rate to detect
2-4 days after identifying garden wheat harvest around, catches small brown rice planthopper more than 500, chooses at random 100, detects small brown rice planthopper colony be with malicious situation according to SN/T1666-2005, calculates the band poison rate of small brown rice planthopper colony;
5) the effective small brown rice planthopper density of inoculation and be with malicious rate requirement
The effectively small brown rice planthopper density of inoculation and be with malicious rate relation to calculate by formula (1):
D＝6×10
6/PVS………………………………………………(1)
In formula:
D---the effectively small brown rice planthopper density of inoculation, unit is a per hectare (head/hm
2);
PVS---small brown rice planthopper is with malicious rate, and unit is percentage (%);
Identify that in garden, small brown rice planthopper density should be lower than 1.5 × 10
8head/hm
2, known according to formula (1) when small brown rice planthopper is with malicious rate to reach 100%, identify in garden small brown rice planthopper density minimum be 6 × 10
6head/hm
2;
Identify that in garden, small brown rice planthopper is with malicious rate to be not less than 4%;
6) management
Identify garden and around in 10m field do not use any insecticide and prevent and treat viral agent, crop straw burning not after wheat harvest; Field fertilizer application, water use management are produced unanimously with conventional, do not use any insecticide and prevent and treat viral agent;
7) investigation period
Identify in garden and grow to while tillering peak period and carry out state of an illness investigation after shoot transplanting equipment, North Jiangsu Area, Jiangsu is in late July;
8) investigation method
Identify after shoot transplanting equipment, to grow in garden and while tillering peak period, add up the incidence of disease:
Now disease plant shows obvious symptom: rice strain seriously stunt not as good as normal plant height 1/3, lobus cardiacus break through inferior lobe leaf sheath and go out, part plant show as from lower pulvinus mouth stretch out in the shape of a spiral, to there is revolving shape in the top of partial blade curling;
And healthy rice strain performance plant height is normal, to tiller vigorous, vitality is strong; Morbidity percentage mean value using 2 repetitions in each test site is added up as the anti-black streak dwarf phenotypic value of kind;
9) division of black streaked dwarf virus of rice resistance rank:
Immunity: the incidence of disease is 0;
High resistance: incidence of disease 0-5%;
Disease-resistant: incidence of disease 5%-10%;
In anti-: incidence of disease 11-20%;
Susceptible: incidence of disease 20%-50%;
High sense: the incidence of disease is higher than 50%.
Beneficial effect
This method can disposablely be carried out anti-black streak dwarf evaluation to a large amount of rice varieties, overcomes the shortcoming that laboratory manually identifies that scale is less, identification of species number is less.This authentication method can be applied to the various fields such as the anti-black streak dwarf rice varieties of screening, varietal resistance evaluation, resistant variety seed selection and the research of genetics of resistance rule.
The inventive method is for more than 1000 rice varieties black streak dwarf Resistance Identifications, and part variety identification result is as shown in table 3.
Accompanying drawing explanation
Fig. 1 rice tillering peak period black streak dwarf feature
A: the disease plant B after black-streaked dwarf virus infects: the morbidity family after black-streaked dwarf virus infects
Specific implementation method
(1) disposable a large amount of rice varieties are carried out to anti-black streak dwarf evaluation
1, identify garden selection
Select the continuous field of retransmitting above black streaked dwarf virus of rice for 3 years, and susceptible check variety of prior year is greater than 40% as identifying garden not preventing and treating under condition the incidence of disease, identifies that garden surrounding plantation wheat entrusts one's child to the care of sb. district as small brown rice planthopper, to guarantee to obtain enough worms source.
2, broadcast cultivation mode
Participate in the variety seeds (containing disease-resistant contrast and susceptible check variety) of evaluation, sowing in 15-20 days before wheat harvest, each material to be identified is broadcast 150 dry seedses, uniform broadcasting 1 row, the long 50cm of row, line-spacing 10cm.Sowing finishes rear field immersion 1-2 days, normal management after emerging, not with medicament insect protected between nursery period.
Length of time rice seedlings grow 5-6 transplants during leaf, and single seedling is planted, distance between rows and hills 15cm × 20cm, and each kind and family are planted 100 caves.Conventional rich water quality management, any agricultural chemicals is not used in land for growing field crops.Twice repetition.
3, small brown rice planthopper density and be with the investigation of malicious rate
1) small brown rice planthopper density survey
For small brown rice planthopper moves to the high-incidence season of identifying garden from wheatland, average, consistent in order to make to identify the small brown rice planthopper density of each point in garden before and after wheat harvest, every mornings 94 of point, afternoons are driven the small brown rice planthopper of identifying in garden with bamboo pole.
2-4 days after the wheat harvest around of evaluation garden, identifies that in garden, small brown rice planthopper density reaches maximum, and therefore selecting this period is the best opportunity of investigation small brown rice planthopper density.Whole evaluation garden is adopted to 5 samplings of diagonal method, and 0.1(0.5 × 0.2 is looked in every some bat) m
2; The moistening rectangle of water (35cm × 50cm) white enamel dish inwall, aims at rice seedling middle and lower part, continuous shooting three times, and each bat looked into the rear white enamel dish that cleans of counting, carries out clapping next time and looks into.
2) small brown rice planthopper is with malicious rate to detect
2-4 days after identifying garden wheat harvest around, catches small brown rice planthopper more than 500, chooses at random 100, detects small brown rice planthopper colony be with malicious situation according to SN/T1666-2005, calculates the band poison rate of small brown rice planthopper colony.
4, the effective small brown rice planthopper density of inoculation and be with malicious rate requirement
The effectively small brown rice planthopper density of inoculation and be with malicious rate relation to calculate by formula (1):
D＝6×10
6/PVS………………………………………………(1)
In formula:
D---the effectively small brown rice planthopper density of inoculation, unit is a per hectare (head/hm
2);
PVS---small brown rice planthopper is with malicious rate, and unit is percentage (%).
According to seeding quantity and sowing spacing in the rows and the line-spacing of identifying garden, per hectare identifies that garden seedling number is about 3 × 10
7strain, every strain rice shoot worm amount is multiplied by small brown rice planthopper to be with malicious rate to approach at 1 o'clock just to reach inoculation requirement, sets every small brown rice planthopper and between the different plants of field, can pass poison 5 times, therefore the effective small brown rice planthopper density of inoculation and to be with malicious rate to close be D=(3 × 10
7÷ 5)/PVS, be formula (1).
1) the small brown rice planthopper density requirements of effectively inoculating
When identifying that in garden, small brown rice planthopper density reaches 1.5 × 10
8head/hm
2time, rice shoot is subject to insect pest heavier, occurs a large amount of dead seedlings, therefore identifies that in garden, small brown rice planthopper density should be lower than 1.5 × 10
8head/hm
2.Known according to formula (1) when small brown rice planthopper is with malicious rate to reach 100%, identify in garden small brown rice planthopper density minimum be 6 × 10
6head/hm
2.
2) effectively the small brown rice planthopper of inoculation is with malicious rate requirement
Identify that in garden, small brown rice planthopper density should be lower than 1.5 × 10
8head/hm
2, under this condition, according to formula (1), calculate, identify that in garden, small brown rice planthopper is with malicious rate to be not less than 4%.
5, district's management is entrusted one's child to the care of sb. in evaluation garden
Identify garden and around in 10m field do not use any insecticide and prevent and treat viral agent, crop straw burning not after wheat harvest.
6, investigation period
Identify in garden and grow to while tillering peak period and carry out state of an illness investigation after shoot transplanting equipment, North Jiangsu Area, Jiangsu is conventionally in late July.
7, investigation method
Identify after shoot transplanting equipment, to grow in garden and while tillering peak period, add up the incidence of disease.Now disease plant shows obvious symptom (list of references: Liu Wancai etc., vigilant southern rice black-streaked dwarf disease is broken out and is caused serious harm, < < China plant protection guide > >, 2010,30 (3): 17-18.):
Rice strain seriously stunt not as good as normal plant height 1/3, lobus cardiacus break through inferior lobe leaf sheath and go out, part plant show as from lower pulvinus mouth stretch out in the shape of a spiral, to there is revolving shape in the top of partial blade curling;
And healthy rice strain performance plant height is normal, tiller vigorous, vitality strong (Fig. 1).In addition, disease plant now seldom has osculant symptom, using the morbidity percentage mean value of 2 repetitions in each test site, as the anti-black streak dwarf phenotypic value of kind, adds up.
8, field rich water quality management
Field fertilizer application, water use management are produced unanimously with conventional, do not use any insecticide and prevent and treat viral agent.
9, the division of black streaked dwarf virus of rice resistance rank:
Immunity: the incidence of disease is 0; High resistance: incidence of disease 0-5%; Anti-: incidence of disease 5%-10%; In anti-: incidence of disease 11-20%; Susceptible: the incidence of disease is higher than 20%, and height is felt.
(2) the effective small brown rice planthopper density of inoculation and be with malicious rate to require experimental basis
Be with under malicious rate 3%, 5% and 8% condition, when connecing worm amount and rising to 8,000,000/mu from 6,000,000/mu, high sense black streaked dwarf virus of rice kind Wu-Yu-Geng 3 incidence of disease ascensional range maximum, is respectively 15%, 10% and 16%; When connecing worm amount and rising to 1,000 ten thousand/mu from 8,000,000/mu, Wu-Yu-Geng 3 death rate ascensional range maximum, be respectively 13%, 12% and 18%(table 2).In addition, in small brown rice planthopper worm amount, be under 8,000,000/mu of conditions, when small brown rice planthopper is with malicious rate to be 3%, the Wu-Yu-Geng 3 incidence of disease is 26%, falls ill lighter; And while being with malicious rate to be respectively 5% and 8%, the incidence of disease has reached 48% and 64%, so be with malicious rate should be not less than 5%.
Table 1 worm amount, with the relation between malicious rate, the incidence of disease and death rate
(3) experimental basis of the each rank evaluation criteria of resistance
Wu-Yu-Geng 3 is under different onset rate condition, and production loss rate, all lower than the incidence of disease, shows that healthy tree exists certain compensating action.When the incidence of disease is 0-5.25%, production loss rate is lower than 1%; Incidence of disease 6.25%-10.5%, production loss 0.8-3.4%; Incidence of disease 12.25-19.5%, production loss 4.1-8.2%; Incidence of disease 23.25%-62%, production loss 10.1-39.4%.
The impact of the different black streak dwarf incidences of disease of table 2 on output
(4) the inventive method is for more than 1000 rice varieties black streak dwarf Resistance Identifications, and part variety identification result is as shown in table 3.
The resistance rank of 112 rice varieties in table 3 Jiangsu and Yunnan Province to black streak dwarf
Claims (1)
1. the Field inoculation authentication method of rice varieties to black streaked dwarf virus of rice resistance, is characterized in that:
1) identify garden selection:
Select the continuous field of retransmitting above black streaked dwarf virus of rice for 3 years, and susceptible check variety of prior year is greater than 40% not preventing and treating the incidence of disease under condition, and after wheat harvest, identifies that in garden, passing virus mediator small brown rice planthopper is with malicious rate higher than 4%, insect density is up to 1.5 × 10
8head/hm
2, minimum is 6 × 10
6head/hm
2field as identifying garden, identify that garden surrounding plantation wheat entrusts one's child to the care of sb. district as small brown rice planthopper, to guarantee to obtain enough worms source;
2) broadcast cultivation mode
Participate in the variety seeds of evaluation, sowing in 15-20 days before wheat harvest, each material to be identified is broadcast 150 dry seedses, uniform broadcasting 1 row, the long 50cm of row, line-spacing 10cm; Sowing finishes rear field immersion 1-2 days, normal management after emerging, not with medicament insect protected between nursery period;
Length of time rice seedlings grow 5-6 transplants during leaf, and single seedling is planted, distance between rows and hills 15cm × 20cm, and each kind and family are planted 100 caves, conventional rich water quality management, any agricultural chemicals is not used in land for growing field crops, twice repetition;
3) small brown rice planthopper density
For small brown rice planthopper moves to the high-incidence season of identifying garden from wheatland, average, consistent in order to make to identify the small brown rice planthopper density of each point in garden before and after wheat harvest, every mornings 94 of point, afternoons are driven the small brown rice planthopper of identifying in garden with bamboo pole;
After identifying garden wheat harvest around, within 2-4 days, investigate small brown rice planthopper density: whole evaluation garden is adopted to 5 samplings of diagonal method, and 0.5m × 0.2m is looked in every some bat; Water moistening rectangle 35cm × 50cm white enamel dish inwall, aims at rice seedling middle and lower part, continuous shooting three times, and each bat looked into the rear white enamel dish that cleans of counting, carries out clapping next time and looks into;
4) small brown rice planthopper is with malicious rate to detect
2-4 days after identifying garden wheat harvest around, catches small brown rice planthopper more than 500, chooses at random 100, detects small brown rice planthopper colony be with malicious situation according to SN/T 1666-2005, calculates the band poison rate of small brown rice planthopper colony;
5) the effective small brown rice planthopper density of inoculation and be with malicious rate requirement
The effectively small brown rice planthopper density of inoculation and be with malicious rate relation to calculate by formula (1):
D＝6×10
6/
PVS?………………………………………………(1)
In formula:
d---the effectively small brown rice planthopper density of inoculation, unit is a per hectare (head/hm
2);
pVS---small brown rice planthopper is with malicious rate, and unit is percentage (%);
Identify that in garden, small brown rice planthopper density should be lower than 1.5 × 10
8head/hm
2, known according to formula (1) when small brown rice planthopper is with malicious rate to reach 100%, identify in garden small brown rice planthopper density minimum be 6 × 10
6head/hm
2;
Identify that in garden, small brown rice planthopper is with malicious rate to be not less than 4%;
6) management
Identify garden and around in 10m field do not use any insecticide and prevent and treat viral agent, crop straw burning not after wheat harvest; Field fertilizer application, water use management are produced unanimously with conventional, do not use any insecticide and prevent and treat viral agent;
7) investigation period
Identify in garden and grow to while tillering peak period and carry out state of an illness investigation after shoot transplanting equipment, North Jiangsu Area, Jiangsu is in late July;
8) investigation method
Identify after shoot transplanting equipment, to grow in garden and while tillering peak period, add up the incidence of disease:
Now disease plant shows obvious symptom: rice strain seriously stunt not as good as normal plant height 1/3, lobus cardiacus break through inferior lobe leaf sheath and go out, part plant show as from lower pulvinus mouth stretch out in the shape of a spiral, to there is revolving shape in the top of partial blade curling;
And healthy rice strain performance plant height is normal, to tiller vigorous, vitality is strong; Morbidity percentage mean value using 2 repetitions in each test site is added up as the anti-black streak dwarf phenotypic value of kind;
9) division of black streaked dwarf virus of rice resistance rank:
Immunity: the incidence of disease is 0;
High resistance: incidence of disease 0-5%;
Disease-resistant: incidence of disease 5%-10%;
In anti-: incidence of disease 11-20%;
Susceptible: incidence of disease 20%-50%;
High sense: the incidence of disease is higher than 50%.
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