CN113229092A - Breeding method for improving resistance to flue-cured tobacco bacterial wilt - Google Patents

Breeding method for improving resistance to flue-cured tobacco bacterial wilt Download PDF

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CN113229092A
CN113229092A CN202110474573.8A CN202110474573A CN113229092A CN 113229092 A CN113229092 A CN 113229092A CN 202110474573 A CN202110474573 A CN 202110474573A CN 113229092 A CN113229092 A CN 113229092A
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bacterial wilt
resistance
disease
flue
cured tobacco
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张振臣
陈俊标
潘晓英
赵伟才
黄跃鹏
袁清华
陈博文
曾涛
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NANXIONG SCIENTIFIC RESEARCH INSTITUTE OF GUANGDONG TOBACCO
CROP Research Institute of Guangdong Academy of Agricultural Sciences
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NANXIONG SCIENTIFIC RESEARCH INSTITUTE OF GUANGDONG TOBACCO
CROP Research Institute of Guangdong Academy of Agricultural Sciences
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G22/00—Cultivation of specific crops or plants not otherwise provided for
    • A01G22/45—Tobacco
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
    • A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
    • A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
    • A01G24/22—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing plant material
    • A01G24/25—Dry fruit hulls or husks, e.g. chaff or coir
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
    • A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
    • A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection

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  • Chemical & Material Sciences (AREA)
  • Inorganic Chemistry (AREA)
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  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The invention provides a breeding method for improving resistance of flue-cured tobacco bacterial wilt, which comprises the following steps: (1) selecting a tobacco variety with high bacterial wilt resistance as a disease-resistant parent, selecting a high-quality flue-cured tobacco variety as a parent to be improved, carrying out hybridization to obtain F1 seeds, planting F1 plants in a field, bagging and selfing to obtain F2 seeds; (2) adopting a certain ralstonia solanacearum concentration as bacterial wilt resistance screening pressure to process the hybrid offspring seedlings, and transplanting the seedlings which do not suffer from bacterial wilt to a bacterial wilt disease garden for field planting after 15-30 days; (3) screening F2-generation individual plants with excellent agronomic characters and without bacterial wilt symptoms in a bacterial wilt disease garden, bagging the individual plants, and selfing to obtain F3-generation seeds; (4) and (4) repeating the steps (2) and (3) until F6, and obtaining the high-generation bacterial wilt-resistant flue-cured tobacco strain. The method selects proper tobacco parents to carry out cross breeding, and obtains the new bacterial wilt-resistant flue-cured tobacco strain through disease pressure and breeding target character screening.

Description

Breeding method for improving resistance to flue-cured tobacco bacterial wilt
Technical Field
The invention particularly relates to a breeding method for improving resistance of flue-cured tobacco bacterial wilt.
Background
Flue-cured tobacco is a commercial crop widely planted in China, is a main raw material crop for cigarette production, is also an important commercial crop, and is one of important factors influencing the quality of tobacco leaves, wherein the variety is the basis of the production of high-quality tobacco leaf raw materials. The excellent variety plays an important role in improving the quality of tobacco leaves, increasing the benefit of unit area, resisting the diseases and insect pests of the tobacco, resisting adverse environmental factors and the like. Therefore, the quality of the flue-cured tobacco variety has great social and economic benefits.
Tobacco bacterial wilt is a bacterial soil-borne disease caused by ralstonia solanacearum, is widely distributed in tropical, subtropical and some temperate regions, and is one of the main diseases threatening the world tobacco production. Tobacco bacterial wilt is a typical vascular bundle disease, all parts of roots, stems and leaves can be damaged, and the most typical symptom is blight. Aiming at the control technology of tobacco bacterial wilt, Chinese scholars at home and abroad research on aspects of chemical, biological and agricultural measures and the like, and all obtain certain control effect. The method takes the cultivation of disease-resistant varieties as the best method, is environment-friendly, has no pollution to soil, and has no problems of pesticide residue exceeding standard and the like.
The following 3 types of tobacco bacterial wilt resistant sources are found in foreign reports: (1) the common tobacco variety T.I.448A collected from Columbia in the United states has higher-level resistance which is polygenic inheritance; (2) turkish aromatic tobacco species Xanthi, having low level resistance, controlled by a part of dominant gene Rxa; (3) japanese local air-cured tobacco species Kokubu, Hatano, Odaruma and Awa, the resistance is controlled by partial dominant gene Rps, and the resistance is from low resistance to medium resistance when the disease pressure is low; the resistance of the local sun-cured tobacco species Enshu, Hatanodaruma and the like in Japan is controlled by a part of dominant genes Rps and polygenes, and the local sun-cured tobacco species have higher-level resistance. T.I.448A is the most widely applied antigen in flue-cured tobacco breeding for disease resistance, and DB101 bred by the method and Coker139, NC95 and Coker319 derived from the DB101 are main parents for breeding for bacterial wilt resistance. When 79-X is used as a source of resistance in flue-cured tobacco breeding, resistant progeny individuals tend to have poor quality leaflets. The resistance of Japanese local air-cured tobacco type is closely linked with poor baking performance, and is not applied to flue-cured tobacco breeding for disease resistance. Zhang Zhenchen et al screened high bacterial wilt resistance germplasm resource GDSY-1 from yellow tobacco sun-cured in Guangdong, which has bacterial wilt resistance superior to DB101 and resistance inheritance different from other resistance sources.
Although the flue-cured tobacco varieties bred by taking T.I.448A as an antigen are more, the varieties with strong resistance are poorer in economic character, and the varieties with better economic character are poorer in resistance, so that the method is a main reason for lacking available bacterial wilt resistant flue-cured tobacco varieties in tobacco production.
Disclosure of Invention
The invention aims to provide a breeding method for improving the resistance of the bacterial wilt of flue-cured tobacco, and solves the technical problem of low resistance of the bacterial wilt of the existing flue-cured tobacco variety.
The purpose of the invention is realized by the following technical scheme:
a breeding method for improving resistance to flue-cured tobacco bacterial wilt comprises the following steps:
(1) selecting tobacco varieties (lines) with high bacterial wilt resistance as disease-resistant parents, selecting high-quality flue-cured tobacco varieties as parents to be improved, hybridizing to obtain F1 seeds, planting F1 plants in a field, bagging and selfing to obtain F2 seeds;
(2) f2 seeds are sowed in a seedling tray, when the seedlings are about 40 days old, a bacterial liquid of ralstonia solanacearum with a certain concentration is poured into the roots of the seedlings or a seedling pool, and after 15-30 days, the seedlings without the bacterial wilt are transplanted to a bacterial wilt disease garden for field planting;
(3) in a bacterial wilt disease garden, when tobacco plants grow to reach the bud stage to the full-bloom stage, eliminating susceptible and early flowering plants, screening F2 single plants with excellent agronomic characters and without bacterial wilt symptoms, bagging the single plants for selfing to obtain F3 generation seeds;
(4) sowing the F3 generation seeds obtained in the step (3) on a seedling raising plate, processing according to the step (2), after bacterial wilt resistance screening, transplanting seedlings which do not suffer from bacterial wilt to a bacterial wilt disease garden for field planting, screening F3 single plants which have excellent agronomic characters and do not have bacterial wilt symptoms, and bagging and selfing the single plants to obtain F4 generation seeds; and repeating the operations of seed treatment, field planting, screening of single plants and bagging of the single plants for selfing until F6 generation seeds are obtained, namely the high-generation bacterial wilt-resistant flue-cured tobacco strain.
In the invention, the disease-resistant parent is 'GDSY-1' or a filial generation taking 'GDSY-1' as a parent.
In the invention, the bacterial liquid concentration of ralstonia solanacearum is 1 × 106～1×109CFU/mL。
Further, the bacterial liquid concentration of the ralstonia solanacearum is obtained as a bacterial wilt resistance screening pressure in the following manner: adopting ralstonia solanacearum with gradient concentration to process disease-resistant parent and susceptible variety seedlings, and selecting ralstonia solanacearum with disease-susceptible variety disease-developing rate more than 90% and disease-developing rate 0< disease-resistant parent disease-developing rate < 20% as bacterial wilt resistance screening pressure.
Further, the gradient concentration of ralstonia solanacearum is 1 × 106CFU/mL、1×107CFU/mL、1×108CFU/mL、1×109CFU/mL。
Furthermore, the susceptible variety is a tobacco variety with high bacterial wilt sensitivity, and the tobacco variety with high bacterial wilt sensitivity is one of Changqiu yellow, Honghuadajinyuan and Cuibi I.
In the process of raising seedlings of hybrid offspring, the seedling raising substrate is prepared by peat soil, carbonized chaff, perlite and loess according to a certain proportion, and the pH value is 4.5-6.5. Under the formula of the seedling culture medium, tobacco seedlings can grow normally, the activity of ralstonia solanacearum is high, and the bacterial wilt resistance screening is facilitated.
Further, the volume ratio of peat soil, carbonized chaff, perlite and loess in the seedling substrate is 2: 1: 1:2-4.
Further, after the seedling is inoculated with the ralstonia solanacearum, the humidity of the matrix is kept between 70 and 90 percent, and the environmental temperature is kept between 25 and 35 ℃. Under set humidity and temperature conditions, ralstonia solanacearum has optimal activity.
In the invention, the standard of the bacterial wilt disease nursery is as follows: the disease incidence is uniform when planting bacterial wilt susceptible varieties in the field, and the disease index of the susceptible varieties is more than 90. And (4) further screening tobacco plants with adult-plant resistance by planting hybrid progeny in a bacterial wilt disease garden.
In the invention, the individual plant of the filial generation with excellent agronomic characters has the following indexes: the plant type is tower type, olive type or cylinder type, the total number of leaves is 20-30, the shape of the leaves is oval or oblong, no petiole exists, the length of the waist leaf is more than 50 cm, and the leaves are layered and yellow.
Compared with the prior art, the invention has the following beneficial effects:
(1) the method for breeding the resistance of the flue-cured tobacco to the bacterial wilt selects proper tobacco parents to carry out cross breeding, adopts the concentration of the ralstonia solanacearum as the screening pressure of the bacterial wilt resistance to process the hybrid offspring, and obtains the new strain of the flue-cured tobacco with the bacterial wilt resistance by screening the bacterial wilt resistance and the agronomic characters of the hybrid offspring in the adult stage.
(2) The invention improves the bacterial wilt resistance of the flue-cured tobacco according to the resistance characteristics, resistance inheritance and biological character characteristics of GDSY-1 varieties, and simultaneously adopts a series of technical measures which are beneficial to the occurrence of the bacterial wilt: the method has the advantages that the method reduces the field workload and the planting scale and improves the breeding efficiency by 3-5 times through screening the bacterial wilt resistance in the seedling stage, and compared with the original breeding method, the method has the characteristics of stronger pertinence, higher efficiency and suitability for breeding new varieties of high-quality and high-bacterial-wilt-resistant flue-cured tobaccos.
(3) According to the invention, the breeding process is optimized according to the resistance inheritance and the agronomic character characteristics of GDSY-1, and the breeding efficiency is greatly improved by selecting a proper bacterial wilt resistance screening pressure and an appropriate agronomic character screening index.
(4) The invention integrates the resistance, quality and yield of the flue-cured tobacco through breeding, has high economic value, can meet the production requirement of the flue-cured tobacco and can be popularized in a large range.
Detailed Description
The present invention is further described below in conjunction with specific examples to better understand and implement the technical solutions of the present invention for those skilled in the art.
Example 1
A breeding method for improving resistance to flue-cured tobacco bacterial wilt comprises the following steps:
(1) selecting a high-quality flue-cured tobacco variety K326 as a female parent and a sun-cured tobacco variety GDSY-1 as a male parent in spring, and carrying out artificial hybrid pollination to obtain F1 generation seeds; f1 plants are planted in the field in autumn, and are bagged and selfed to obtain F2 seeds.
(2) With a gradient concentration of 1X 106CFU/mL、1×107CFU/mL、1×108CFU/mL、1×109Treating disease-resistant parent sun-cured tobacco variety GDSY-1 and susceptible variety Changneck yellow seedling (seedling age 40 days) with ralstonia solanacearum in CFU/mL, and selecting the susceptible variety with diseased plant rate>90% and 0<Disease-resistant parent disease incidence<The bacterial wilt concentration of 20 percent is used as the bacterial wilt resistance screening pressure, and the bacterial wilt concentration of the bacterial wilt resistance screening pressure is finally determined to be 1 multiplied by 107CFU/mL。
(3) The concentration is 1 x 107Pouring the bacterial liquid of the ralstonia solanacearum of CFU/mL into a seedling pool of 10000F 2 seedlings (about 40 days old), wherein the seedling matrix adopts peat soil, carbonized chaff, perlite and loess according to the volume ratio of 2: 1: 1:4, the pH value of the seedling substrate is 4.5, the humidity of the seedling substrate is kept at 70-80%, the environmental temperature is 25-30 ℃, and 2000 seedlings without disease symptoms are transplanted to a bacterial wilt disease garden after 20 days to continue to be planted in a field.
(4) In a bacterial wilt disease garden, when tobacco plants grow to reach the bud stage to the full-bloom stage, infected and early-flowering plants are eliminated, 25 single plants of F2 generation with excellent agronomic characters and without bacterial wilt symptoms are screened, and the single plants are bagged and selfed to obtain seeds of F3 generation.
(5) And (3) sowing 200 seeds of each of the 25F 3 generations obtained in the step (4) on a seedling raising tray, after bacterial wilt resistance screening, transplanting 2400 seedlings which do not have bacterial wilt into a bacterial wilt disease garden for field planting, screening 20F 3 generations which have excellent agronomic characters and do not have bacterial wilt symptoms, and bagging and selfing the individual plants to obtain F4 generation seeds.
(6) And (3) sowing 200 seeds of each of the 20F 4 generations obtained in the step (5) on a seedling raising plate, after bacterial wilt resistance screening, transplanting 2000 seedlings which do not have bacterial wilt into a bacterial wilt disease garden for field planting, screening 10F 4 generations which have excellent agronomic characters and do not have bacterial wilt symptoms, and bagging and selfing the individual plants to obtain F5 generation seeds.
(7) And (3) sowing 200 seeds of each of the 10F 5 generations obtained in the step (6) on a seedling raising plate, after bacterial wilt resistance screening, transplanting 500 seedlings which do not have bacterial wilt into a bacterial wilt disease garden to carry out field planting, screening 5 single plants of the F5 generation which have excellent agronomic characters and do not have bacterial wilt symptoms, bagging and selfing the single plants to obtain F6 generation seeds which are respectively named as Qing-1, Qing-2, Qing-3, Qing-4 and Qing-5.
Example 2
A breeding method for improving resistance to flue-cured tobacco bacterial wilt comprises the following steps:
(1) selecting a high-quality flue-cured tobacco variety K326 as a female parent in spring, and taking a hybridization progeny 'green-2' of the K326 and GDSY-1 as a male parent to carry out artificial hybridization pollination to obtain F1 generation seeds; planting F1 plant in field in autumn, bagging and selfing to obtain F2 generation seeds;
(2) adopting the ralstonia solanacearum concentration of 1 × 106And the CFU/mL is used as a bacterial wilt resistance screening and pressure treatment method for 8000F 2 seedlings (about 40 days old), and the seedling culture substrate adopts peat soil, carbonized chaff, perlite and loess according to the volume ratio of 2: 1: 1:2, the pH value of the seedling substrate is 6.5, the humidity of the seedling substrate is 70%, the seedling temperature is 25-30 ℃, and after 28 days, 1000 seedlings without bacterial wilt are transplanted to a bacterial wilt disease garden for field planting.
(3) In a bacterial wilt disease garden, when tobacco plants grow to reach the bud stage to the full-bloom stage, infected and early-flowering plants are eliminated, 20 single plants of F2 generation with excellent agronomic characters and without bacterial wilt symptoms are screened, and the single plants are bagged and selfed to obtain seeds of F3 generation.
(4) And (3) sowing 200 seeds of each of the 20F 3 generations obtained in the step (3) on a seedling raising plate, after bacterial wilt resistance screening, transplanting 1200 seedlings which do not have bacterial wilt into a bacterial wilt disease garden for field planting, screening 18 single plants of the F3 generation which have excellent agronomic characters and do not have bacterial wilt symptoms, and bagging and selfing the single plants to obtain F4 generation seeds.
(5) And (3) sowing 200 seeds of each of 18F 4 generations obtained in the step (4) on a seedling raising plate, after bacterial wilt resistance screening, transplanting 1000 seedlings without bacterial wilt into a bacterial wilt disease garden for field planting, screening 9 single plants of F4 generations with excellent agronomic characters and without bacterial wilt symptoms, and bagging and selfing the single plants to obtain F5 generation seeds.
(6) And (3) sowing 200 seeds of each of the 9F 5 generations obtained in the step (5) on a seedling raising plate, after bacterial wilt resistance screening, transplanting 400 seedlings which do not have bacterial wilt into a bacterial wilt disease garden for field planting, screening 6 single plants of the F5 generations which have excellent agronomic characters and do not have bacterial wilt symptoms, and bagging and selfing the single plants to obtain F6 generation seeds.
(7) And (3) performing variety comparison test and bacterial wilt resistance identification on the 6 strains obtained in the step (6), repeating the steps for three times, wherein each cell is 0.05 mu, performing economic character investigation on the flue-cured tobacco, integrating the yield, the output value and the resistance, and selecting new flue-cured tobacco strains 'green-2-2' and 'green-2-5' with high yield, high output value and strong resistance from the flue-cured tobacco strains, wherein the character questionnaire is shown in table 1.
TABLE 1 survey of traits of new lines "Qing-2-2" and "Qing-2-5" with high resistance to bacterial wilt
Figure BDA0003046955250000071
Note: the identification of bacterial wilt resistance is carried out in disease nursery; the economic character investigation test field has no bacterial wilt; yellow and DB101 are control varieties for resistance identification, and K326 is a control variety for resistance identification and economic trait survey.
It can be known that the average yield per mu of the green-2-2 reaches 158.45 kg, the yield per mu value reaches 4705.74 yuan, the average price reaches 29.60 yuan/kg, the upper grade tobacco proportion is 79.06%, the middle grade tobacco proportion is 98.21%, and the amplification is respectively compared with a reference variety K326: 11.19%, 19.86%, 7.44%, 21.80% and 2.92%. The resistance identification result shows that the 'Qing-2-2' has high resistance to tobacco bacterial wilt, and the resistance is superior to 'DB 101' and 'K326'.
The average yield per mu of green-2-5 reaches 158.31 kg, the yield per mu value is 4441.92 yuan, the average price is 28.06 yuan/kg, the upper grade tobacco proportion is 81.69%, the middle grade tobacco proportion is 96.24%, and the amplification is respectively increased compared with a reference variety K326: 11.09%, 13.14%, 1.85%, 25.85% and 0.86%. The resistance identification result shows that the 'Qing-2-2' has high resistance to tobacco bacterial wilt, and the resistance is superior to 'DB 101' and 'K326'.
Example 3
A breeding method for improving resistance to flue-cured tobacco bacterial wilt comprises the following steps:
(1) selecting a high-quality flue-cured tobacco variety Yunyan 87 as a female parent, and taking a hybrid progeny green-3 of K326 and GDSY-1 as a male parent to carry out artificial hybrid pollination to obtain F1 generation seeds; f1 plants are planted in the field, and the F2 generation seeds are obtained by bagging and selfing;
(2) adopting the ralstonia solanacearum concentration of 1 × 108And the CFU/mL is used as a bacterial wilt resistance screening and pressure treatment method for 4000F 2 seedlings (the seedling age is about 40 days), and the seedling culture substrate adopts peat soil, carbonized chaff, perlite and loess according to the volume ratio of 2: 1: 1:3, the pH value of the seedling substrate is 5.5, the humidity of the seedling substrate is 90%, the seedling temperature is 30-35 ℃, and 2000 seedlings without bacterial wilt are transplanted to a bacterial wilt disease garden for field planting after 15 days.
(3) In a bacterial wilt disease garden, when tobacco plants grow to reach the bud stage to the full-bloom stage, infected and early-flowering plants are eliminated, 33 single plants of the F2 generation with excellent agronomic characters and without bacterial wilt symptoms are screened, and the single plants are bagged and selfed to obtain seeds of the F3 generation.
(4) And (3) sowing 200 seeds of each of 33F 3 generations obtained in the step (3) on a seedling raising plate, after bacterial wilt resistance screening, transplanting 2000 seedlings which do not suffer from bacterial wilt to a bacterial wilt disease garden for field planting, screening 20F 3 generations which have excellent agronomic characters and do not have bacterial wilt symptoms, and bagging and selfing the individual plants to obtain F4 generation seeds.
(5) And (3) sowing 200 seeds of each of the 20F 4 generations obtained in the step (4) on a seedling raising plate, after bacterial wilt resistance screening, transplanting 1500 seedlings which do not have bacterial wilt into a bacterial wilt disease garden for field planting, screening 6F 4 generations which have excellent agronomic characters and do not have bacterial wilt symptoms, and bagging and selfing the single plants to obtain F5 generations of seeds.
(6) And (3) sowing 200 seeds of each of the 6F 5 generations obtained in the step (5) on a seedling raising plate, after bacterial wilt resistance screening, transplanting 500 seedlings which do not have bacterial wilt into a bacterial wilt disease garden for field planting, screening 20F 5 generations which have excellent agronomic characters and do not have bacterial wilt symptoms, and bagging and selfing the individual plants to obtain F6 generation seeds.
(7) And (3) performing variety comparison test and bacterial wilt resistance identification on the 20 strains obtained in the step (6), repeating the steps for three times, wherein each cell is 0.05 mu, performing economic character investigation on the flue-cured tobacco, integrating the yield, the output value and the resistance, and selecting new flue-cured tobacco strains 'green-3-11' and 'green-3-15' with high yield, high output value and strong resistance from the flue-cured tobacco strains, wherein the character questionnaire is shown in a table 2.
TABLE 2 survey form of traits for new lines "Qing-3-11" and "Qing-3-15" with high bacterial wilt resistance
Figure BDA0003046955250000091
Note: the identification of bacterial wilt resistance is carried out in disease nursery; the economic character investigation test field has no bacterial wilt; yellow and DB101 are control varieties for resistance identification, and K326 is a control variety for resistance identification and economic trait survey.
It can be known that the average yield per mu of the green-3-11 reaches 187.43 kg, the yield per mu value is 5227.24 yuan, the average price is 27.89 yuan/kg, the upper grade tobacco proportion is 69.88%, the middle grade tobacco proportion is 97.68%, and the amplification is respectively increased compared with a reference variety K326: 31.53%, 33.15%, 1.23%, 7.66% and 2.37%. The resistance identification result shows that the 'Qing-3-11' has high resistance to tobacco bacterial wilt, and the resistance of the 'Qing-3-11' is superior to that of 'DB 101' and 'K326'.
The average yield per mu of green-3-15 reaches 188.33 kg, the yield per mu value is 5414.83 yuan, the average price is 28.75 yuan/kg, the upper grade tobacco proportion is 70.80%, the middle grade tobacco proportion is 99.00%, and the amplification is respectively increased compared with a reference variety K326: 32.16%, 37.93%, 4.36%, 9.07% and 3.75%. The resistance identification result shows that the 'Qing-3-15' has high resistance to tobacco bacterial wilt, and the resistance of the 'Qing-3-15' is superior to that of 'DB 101' and 'K326'.
The above embodiments illustrate various embodiments of the present invention in detail, but the embodiments of the present invention are not limited thereto, and those skilled in the art can achieve the objectives of the present invention based on the disclosure of the present invention, and any modifications and variations based on the concept of the present invention fall within the scope of the present invention, which is defined by the claims.

Claims (9)

1. A breeding method for improving resistance of flue-cured tobacco bacterial wilt is characterized by comprising the following steps:
(1) selecting tobacco varieties with high bacterial wilt resistance as disease-resistant parents, selecting high-quality flue-cured tobacco varieties as parents to be improved, hybridizing to obtain F1 seeds, planting F1 plants in a field, bagging and selfing to obtain F2 seeds;
(2) f2 seeds are sowed in a seedling tray, when the seedlings are 40 days old, the bacterial liquid of ralstonia solanacearum with certain concentration is poured into the roots of the seedlings or a seedling pond, and after 15-30 days, the seedlings without the ralstonia solanacearum are transplanted to a bacterial wilt disease garden for field planting;
(3) in a bacterial wilt disease garden, when tobacco plants grow to reach the bud stage to the full-bloom stage, eliminating susceptible and early flowering plants, screening F2 single plants with excellent agronomic characters and without bacterial wilt symptoms, bagging the single plants for selfing to obtain F3 generation seeds;
(4) sowing the F3 generation seeds obtained in the step (3) on a seedling raising plate, processing according to the step (2), after bacterial wilt resistance screening, transplanting seedlings which do not suffer from bacterial wilt to a bacterial wilt disease garden for field planting, screening F3 single plants which have excellent agronomic characters and do not have bacterial wilt symptoms, and bagging and selfing the single plants to obtain F4 generation seeds; and repeating the operations of seed treatment, field planting, screening of single plants and bagging of the single plants for selfing until F6 generation seeds are obtained, namely the high-generation bacterial wilt-resistant flue-cured tobacco strain.
2. A breeding method for improving resistance to flue-cured tobacco bacterial wilt according to claim 1, wherein the disease-resistant parent is "GDSY-1" or a filial generation with "GDSY-1" as the parent.
3. A breeding method for improving resistance to bacterial wilt of flue-cured tobacco according to claim 1, wherein the bacterial liquid concentration of ralstonia solanacearum is 1X 106～1×109CFU/mL。
4. A breeding method for improving resistance to bacterial wilt of flue-cured tobacco according to claim 3, wherein the bacterial liquid concentration of ralstonia solanacearum is obtained as the screening pressure for resistance to bacterial wilt by: adopting ralstonia solanacearum with gradient concentration to process disease-resistant parent and susceptible variety seedlings, and selecting ralstonia solanacearum with disease-susceptible variety disease-developing rate more than 90% and disease-developing rate 0< disease-resistant parent disease-developing rate < 20% as bacterial wilt resistance screening pressure.
5. A breeding method for improving resistance to flue-cured tobacco bacterial wilt according to claim 4, wherein the susceptible parent is a tobacco variety with high susceptibility to bacterial wilt, and the tobacco variety with high susceptibility to bacterial wilt is one of Changqihuang, honghuadajinyuan or Cuibi I.
6. A breeding method for improving the resistance to the bacterial wilt of flue-cured tobacco as claimed in claim 1, wherein the seedling raising substrate is prepared from peat soil, carbonized chaff, perlite and loess according to a certain proportion, and the pH value of the seedling raising substrate is 4.5-6.5.
7. A breeding method for improving the resistance to the bacterial wilt of flue-cured tobacco as claimed in claim 1, wherein after the seedling is inoculated with the ralstonia solanacearum, the humidity of the substrate is kept at 70-90% and the environmental temperature is kept at 25-35 ℃.
8. A breeding method for improving the resistance to the bacterial wilt of flue-cured tobacco according to claim 1, characterized in that the standard of the disease nursery is as follows: the disease incidence is uniform when planting bacterial wilt susceptible varieties in the field, and the disease index of the susceptible varieties is more than 90.
9. A breeding method for improving resistance to flue-cured tobacco bacterial wilt according to claim 1, wherein the individual hybrid progeny with excellent agronomic characteristics have the following indexes: the plant type is tower type, olive type or cylinder type, the total number of leaves is 20-30, the shape of the leaves is oval or oblong, no petiole exists, the length of the waist leaf is more than 50 cm, and the leaves are layered and yellow.
CN202110474573.8A 2021-04-29 2021-04-29 Breeding method for improving resistance to flue-cured tobacco bacterial wilt Pending CN113229092A (en)

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