阅读说明：本技术 一种无性高产咖啡选育方法 (Asexual high-yield coffee breeding method ) 是由 周峥 李豫 李晓秋 于 2019-12-03 设计创作，主要内容包括：一种无性高产咖啡选育方法,包括以下步骤：1)选择小粒种、中粒种各五个品种的果实,采用固相微萃取技术通过HS-SPME以及GC-MS代谢组学分析挥发性成分；2)选择呋喃类化合物、吡嗪类和醛酮化合物含量高的小粒种、中粒种进行芽接；接芽成活生长2个月后进行二次嫁接,砧木采用直径为0.4～1.4cm的GT1无性系种子实生营养袋育苗,嫁接高度距离地面小于或等于10cm；3)采用专用育苗基质提高根系活力的进行培育8个月；4)8个月之后移栽到田间进行常规的管理并根据咖啡树的需肥特点进行平衡施肥。(A asexual high-yield coffee breeding method comprises the following steps: 1) selecting fruits of five varieties including small-grain variety and medium-grain variety, and analyzing volatile components through HS-SPME and GC-MS metabonomics by adopting a solid-phase microextraction technology; 2) selecting small-grain seeds and medium-grain seeds with high furan compounds, pyrazine compounds and aldehyde ketone compounds for bud grafting; carrying out secondary grafting after the scion bud survives and grows for 2 months, wherein the stock is grown by adopting a GT1 clone seed seedling nutrition bag with the diameter of 0.4-1.4 cm, and the grafting height is less than or equal to 10cm away from the ground; 3) the root system activity is improved by adopting a special seedling raising substrate for cultivation for 8 months; 4) after 8 months, transplanting the coffee tree into the field for conventional management and carrying out balanced fertilization according to the fertilizer requirement characteristics of the coffee tree.)

1. A asexual high-yield coffee breeding method is characterized by comprising the following steps:

1) selecting fruits of five varieties including small-grain variety and medium-grain variety, and analyzing volatile components through HS-SPME and GC-MS metabonomics by adopting a solid-phase microextraction technology;

2) selecting small-grain seeds and medium-grain seeds with high furan compounds, pyrazine compounds and aldehyde ketone compounds for bud grafting; carrying out secondary grafting after the scion bud survives and grows for 2 months, wherein the stock is grown by adopting a GT1 clone seed seedling nutrition bag with the diameter of 0.4-1.4 cm, and the grafting height is less than or equal to 10cm away from the ground;

3) the root system activity is improved by adopting a special seedling raising substrate for cultivation for 8 months;

4) after 8 months, transplanting the coffee tree into the field for conventional management and carrying out balanced fertilization according to the fertilizer requirement characteristics of the coffee tree.

2. The selective breeding method according to claim 1, wherein HS-SPME is performed on coffee bean samples by arrow-shaped solid phase microextraction injection, and then the extract is analyzed by a gas chromatography-mass spectrometer under GC-MS chromatographic conditions: DB-5MS capillary column, carrier gas is high-purity helium, the flow rate is 1.0mL/min, the temperature of a sample inlet is 260 ℃, the sample injection amount is 1 muL, split-flow sample injection is carried out, the split-flow ratio is 6:1, the solvent delay is 5min, and the GC-MS mass spectrum conditions are as follows: and (3) bombarding the ion source by electrons, wherein the temperature of the ion source is 230 ℃, the temperature of the quadrupole is 150 ℃, the electron energy is 70eV, and the mass scanning range m/z is 35-550.

3. The selective breeding method according to claim 2, wherein the HS-SPME further comprises a step of sample pretreatment: taking coffee beans, grinding the coffee beans to obtain coffee bean powder with the particle size of 20-50 meshes, weighing 5g of coffee bean powder in a 20mL headspace extraction bottle, adding 10mL of distilled water, screwing a bottle cap, and sealing.

4. The breeding method according to claim 1, wherein the specific steps of the budding are as follows: cutting or sawing off the stock at a position 0.1-0.2 cm above the grafting position, picking the phloem away from the xylem, cutting the base of the coffee branch into a 1.5-2.0 cm long horse ear-shaped long cutting surface, cutting the opposite tip into a small cutting surface with the length less than 0.3cm, and fixing the small cutting surface with the stock; after cambiums of the stocks and the bud sheets are completely healed, removing binders on two sides of bud points on the bud sheets to expose the bud points, simultaneously keeping 1-2 longest primary branches and leaves thereof on the stocks, completely cutting off the rest primary branches, removing all the binders after the bud points germinate, performing secondary grafting by taking tender tips of the grafted bud seedlings in the sprouting period and the leaf expanding period as scions, and performing grafting by adopting an indoor bark grafting method.

5. The breeding method according to claim 4, wherein the medium-sized seeds are used as rootstocks and the small-sized seeds are used as scions.

6. The breeding method according to claim 1, wherein the special seedling raising substrate is prepared by crushing and mixing coconut shells, Dawang palm fallen leaves and plant ash according to a volume ratio of 3:1:1, adding 5% of quicklime and 10% of shell powder according to the total volume, fully mixing, naturally decomposing, fertilizing for 2-3 times during the period, and decomposing to dark brown to be used as the organic fertilizer for coffee inter-plant grafting seedling raising.

7. The breeding method according to claim 1, wherein the steps of balanced fertilization are as follows: digging an annular shallow ditch at the projection position of the coffee crown in the current year 10-12 months, wherein the ditch depth is 10cm, broadcasting plant ash, the application amount of the plant ash is 0.3-0.5kg, in the next year 10-12 months, collecting a soil sample, drying the soil sample to detect soil nutrients, and combining the nutrients required by the growth of coffee plants in the coffee kingdom according to the mass ratio of N: p₂O₅：K₂O is 1: 0.2-1.5: the required amount of the nitrogenous fertilizer, the phosphate fertilizer and the potash fertilizer is calculated according to the proportion of 0.2-1.5, and the calculation method is that the N content in the nitrogenous fertilizer, the phosphate fertilizer and the potash fertilizer is calculated according to the proportion: p₂O₅：K₂And calculating the content of O, fully and uniformly mixing the three fertilizers to prepare additional fertilizers, ditching two sides of each row of tea trees, applying the phosphorus-potassium fertilizer for the first time in 5 months at intervals of one ditch, applying the phosphorus-potassium fertilizer for the second time in 8 months, and applying the phosphorus-potassium fertilizer for the third time and the fourth time in 3 months and 6 months of the second year respectively, wherein the method is the same as the method for applying the phosphorus-potassium fertilizer for the first time and the second time.

8. The breeding method according to claim 1, wherein the balanced fertilization further comprises foliar topdressing: the boron fertilizer and the zinc fertilizer are prepared into a solution with the concentration of 0.30-0.50% according to the mass ratio of B to Zn to 1 to 3, and the solution is sprayed on the leaf surfaces in 4 months and 8 months every year.

9. The breeding method according to claim 1, wherein the medium-grain coffee is No. 24, No. 24-1, No. 24-10, Xing 27, Xing 28 or Xing 31 medium-grain coffee.

Technical Field

The invention relates to the technical field of agricultural cultivation, in particular to a asexual high-yield coffee breeding method.

Background

Coffee type coffee is a plant of the genus coffee (Coffea) of the family Rubiaceae (Rubiaceae), which is considered by Chevalier research to be 4 groups of 66 plants, generally referred to as coffee of the group Eucof-fea (Coffea-fea) of the species Coffea arabica, the species Coffearobus-ta, the species Coffea (coffealiberiica) and the species Coffeaexcelsa (Coffeaexcelsa), wherein productive cultivation mainly comprises the species Coffea and the species mesocarpa, the species microphyte occupies about 80% of cultivation area, the species mesocarpa occupies 20%, and other species mainly serve as germplasm conservation and breeding materials without large-scale cultivation.

During the growth process of coffee, the coffee is easily affected by adverse factors such as diseases, insect pests, cold damage, drought and the like, and the selective breeding of resistant varieties is an important breeding target of various coffee producing countries. The coffee leaf rust is a main disease of coffee trees, and has the characteristics of rapid outbreak, serious harm and the like, so that the breeding of rust-resistant varieties is always the target of coffee breeders. The grape dental coffee rust research center adopts the HibridodeTimor which is a small-grain and medium-grain natural hybrid with strong rust resistance to carry out artificial hybridization with the high-yield and high-quality Caturra, and breeds a new variety of the Catimer with rust resistance, high yield, short stalk and high quality through multiple backcrosses, thereby solving the problem puzzling the coffee breeding world; however, other resistance breeding is relatively delayed, and research work of insect-resistant breeding, drought-resistant breeding and cold-resistant breeding should be enhanced.

The common breeding methods at present include selective breeding, crossbreeding, mutation breeding (mutation breeding is also divided into physical mutation breeding and chemical mutation breeding), biotechnology breeding, etc., but the breeding methods have the following defects in different degrees: 1. the breeding period is long, the variety breeding from the hybrid rice generally needs several years or even decades, for example, the time of nearly 20 years is spent on breeding a new variety 'bar 1' of the side ginseng from selection, purification, elimination, strain comparison and the like of Xuzhao dynasty royal jelly; 2. the breeding success rate is low; 3. the predictability of breeding is low, and breeders often cultivate dozens or even hundreds of hybrid combinations, but the comprehensive performance of new hybrid lines is poor. Therefore, the search for a new plant breeding method has been urgently awaited.

Disclosure of Invention

In view of the above, the present invention aims to provide a asexual high-yield coffee breeding method, such that the cultivation method can significantly improve the yield and variety of coffee.

In order to achieve the above purpose, the invention provides the following technical scheme: a asexual high-yield coffee breeding method is characterized by comprising the following steps:

1) selecting fruits of five varieties including small-grain variety and medium-grain variety, and analyzing volatile components through HS-SPME and GC-MS metabonomics by adopting a solid-phase microextraction technology;

2) selecting small-grain seeds and medium-grain seeds with high furan compounds, pyrazine compounds and aldehyde ketone compounds for bud grafting; carrying out secondary grafting after the scion bud survives and grows for 2 months, wherein the stock is grown by adopting a GT1 clone seed seedling nutrition bag with the diameter of 0.4-1.4 cm, and the grafting height is less than or equal to 10cm away from the ground;

3) the root system activity is improved by adopting a special seedling raising substrate for cultivation for 8 months;

4) after 8 months, transplanting the coffee tree into the field for conventional management and carrying out balanced fertilization according to the fertilizer requirement characteristics of the coffee tree.

Preferably, HS-SPME is carried out on coffee bean samples by arrow type solid phase microextraction sample introduction, and then the extract is analyzed by a gas chromatography-mass spectrometer, wherein the GC-MS chromatographic conditions are as follows: DB-5MS capillary column, carrier gas is high-purity helium, the flow rate is 1.0mL/min, the temperature of a sample inlet is 260 ℃, the sample injection amount is 1 muL, split-flow sample injection is carried out, the split-flow ratio is 6:1, the solvent delay is 5min, and the GC-MS mass spectrum conditions are as follows: and (3) bombarding the ion source by electrons, wherein the temperature of the ion source is 230 ℃, the temperature of the quadrupole is 150 ℃, the electron energy is 70eV, and the mass scanning range m/z is 35-550.

Preferably, the HS-SPME further comprises the following steps of sample pretreatment: taking coffee beans, grinding the coffee beans to obtain coffee bean powder with the particle size of 20-50 meshes, weighing 5g of coffee bean powder in a 20mL headspace extraction bottle, adding 10mL of distilled water, screwing a bottle cap, and sealing.

Preferably, the budding comprises the following specific steps: cutting or sawing off the stock at a position 0.1-0.2 cm above the grafting position, picking the phloem away from the xylem, cutting the base of the coffee branch into a 1.5-2.0 cm long horse ear-shaped long cutting surface, cutting the opposite tip into a small cutting surface with the length less than 0.3cm, and fixing the small cutting surface with the stock; after cambiums of the stocks and the bud sheets are completely healed, removing binders on two sides of bud points on the bud sheets to expose the bud points, simultaneously keeping 1-2 longest primary branches and leaves thereof on the stocks, completely cutting off the rest primary branches, removing all the binders after the bud points germinate, performing secondary grafting by taking tender tips of the grafted bud seedlings in the sprouting period and the leaf expanding period as scions, and performing grafting by adopting an indoor bark grafting method.

Among them, preferably, medium-sized seeds are used as rootstocks, and small-sized seeds are used as scions.

Preferably, the special seedling raising substrate is prepared by crushing and mixing coconut shells, Dawang palm fallen leaves and plant ash according to the volume ratio of 3:1:1, adding 5% of quicklime and 10% of shell powder according to the total volume, fully mixing uniformly, naturally decomposing, turning over for 2-3 times during the period, and decomposing to dark brown to be used as the organic fertilizer for raising seedlings of coffee interspecies grafting seedlings.

Preferably, the balanced fertilization comprises the following specific steps: digging an annular shallow ditch at the projection position of the coffee crown in the current year 10-12 months, wherein the ditch depth is 10cm, broadcasting plant ash, the application amount of the plant ash is 0.3-0.5kg, in the next year 10-12 months, collecting a soil sample, drying the soil sample to detect soil nutrients, and combining the nutrients required by the growth of coffee plants in the coffee kingdom according to the mass ratio of N: p₂O₅：K₂O is 1: 0.2-1.5: the required amount of the nitrogenous fertilizer, the phosphate fertilizer and the potash fertilizer is calculated according to the proportion of 0.2-1.5, and the calculation method is that the N content in the nitrogenous fertilizer, the phosphate fertilizer and the potash fertilizer is calculated according to the proportion: p₂O₅：K₂And calculating the content of O, fully and uniformly mixing the three fertilizers to prepare additional fertilizers, ditching two sides of each row of tea trees, applying the phosphorus-potassium fertilizer for the first time in 5 months at intervals of one ditch, applying the phosphorus-potassium fertilizer for the second time in 8 months, and applying the phosphorus-potassium fertilizer for the third time and the fourth time in 3 months and 6 months of the second year respectively, wherein the method is the same as the method for applying the phosphorus-potassium fertilizer for the first time and the second time.

Wherein, as preferred, balanced fertilization still includes the foliage dressing: the boron fertilizer and the zinc fertilizer are prepared into a solution with the concentration of 0.30-0.50% according to the mass ratio of B to Zn to 1 to 3, and the solution is sprayed on the leaf surfaces in 4 months and 8 months every year.

Wherein, the medium-grain coffee is preferably No. 24, No. 24-1, No. 24-10, Xing 27, Xing 28 or Xing 31 medium-grain coffee.

Drawings

FIG. 1 is a gas chromatography-mass spectrometry analysis diagram;

FIG. 2 is a diagram of coffee tree grafting;

FIG. 3 shows the third year yield of the coffee tree selected by the present application.

Detailed Description

1) Selecting fruits of five varieties including small-grain variety and medium-grain variety, and analyzing volatile components through HS-SPME and GC-MS metabonomics by adopting a solid-phase microextraction technology;

2) selecting small-grain seeds and medium-grain seeds with high furan compounds, pyrazine compounds and aldehyde ketone compounds for bud grafting; carrying out secondary grafting after the scion bud survives and grows for 2 months, wherein the stock is grown by adopting a GT1 clone seed seedling nutrition bag with the diameter of 0.4-1.4 cm, and the grafting height is less than or equal to 10cm away from the ground;

3) the root system activity is improved by adopting a special seedling raising substrate for cultivation for 8 months;

4) after 8 months, transplanting the coffee tree into the field for conventional management and carrying out balanced fertilization according to the fertilizer requirement characteristics of the coffee tree.

Preferably, HS-SPME is carried out on coffee bean samples by arrow type solid phase microextraction sample introduction, and then the extract is analyzed by a gas chromatography-mass spectrometer, wherein the GC-MS chromatographic conditions are as follows: DB-5MS capillary column, carrier gas is high-purity helium, the flow rate is 1.0mL/min, the temperature of a sample inlet is 260 ℃, the sample injection amount is 1 muL, split-flow sample injection is carried out, the split-flow ratio is 6:1, the solvent delay is 5min, and the GC-MS mass spectrum conditions are as follows: and (3) bombarding the ion source by electrons, wherein the temperature of the ion source is 230 ℃, the temperature of the quadrupole is 150 ℃, the electron energy is 70eV, and the mass scanning range m/z is 35-550.

Preferably, the HS-SPME further comprises the following steps of sample pretreatment: taking coffee beans, grinding the coffee beans to obtain coffee bean powder with the particle size of 20-50 meshes, weighing 5g of coffee bean powder in a 20mL headspace extraction bottle, adding 10mL of distilled water, screwing a bottle cap, and sealing.

Preferably, the budding comprises the following specific steps: the pre-grafted coffee branches are 7-month-old robust and straight branches. Cutting or sawing off the stock at a position 0.1-0.2 cm above the grafting position, picking the phloem away from the xylem, cutting the base of the coffee branch into a 1.5-2.0 cm long horse ear-shaped long cutting surface, cutting the opposite tip into a small cutting surface with the length less than 0.3cm, and fixing the small cutting surface with the stock; after cambiums of the stocks and the bud sheets are completely healed, removing binders on two sides of bud points on the bud sheets to expose the bud points, simultaneously keeping 1-2 longest primary branches and leaves thereof on the stocks, completely cutting off the rest primary branches, removing all the binders after the bud points germinate, performing secondary grafting by taking tender tips of the grafted bud seedlings in the sprouting period and the leaf expanding period as scions, and performing grafting by adopting an indoor bark grafting method.

Coffee has obvious top advantages, and the top advantages can be broken by cutting the anvil, so that scion germination is promoted. The invention keeps partial first-stage branches and leaves which can be used for photosynthesis, provides nutrition for the scions, promotes the rapid growth of the scions, improves the growth vigor and provides shade for young tender scions; the most important thing is that the tree sap flow is kept, the phenomenon of scion death caused by rootstock retrogradation is reduced, meanwhile, the period of cutting out the rootstock is improved, the scion germinates to 10-20cm, at the moment, the scion has 1-2 pairs of first-level branches, and 3-4 pairs of leaves, and can perform photosynthesis to provide nutrition for the scion, and meanwhile, the tree sap flow is kept, the rootstock is not easy to generate the retrogradation phenomenon, and therefore the defects that the bud grafting germination rate is low and the bud grafting branches are poor in growth vigor in the traditional method are overcome. Compared with the traditional method, the method for bud grafting of coffee is higher in bud grafting germination rate. Meanwhile, the growth vigor of the bud grafting branch in the aspects of stem thickness, stem length, crown breadth, primary plant division number and leaf number is also better.

The stock shearing is carried out twice, and the first-level branches and the leaves thereof are selectively reserved between the two stock shearing, so that the defects of low bud grafting germination rate and poor growth of bud grafting branches of the traditional method by one stock shearing are overcome.

Among them, preferably, medium-sized seeds are used as rootstocks, and small-sized seeds are used as scions.

The special seedling raising matrix is prepared by crushing and mixing coconut shells, palm fallen leaves of king palm and plant ash according to the volume ratio of 3:1:1, adding 5% of quicklime and 10% of shell powder according to the total volume, fully mixing, naturally decomposing, turning over the fertilizer for 2-3 times during the period, and decomposing to black brown to be used as the organic fertilizer for raising seedlings of coffee inter-graft seedlings.

Preferably, digging planting holes before field planting after grafting of young coffee plants, airing soil dug out of the planting holes, returning surface soil to the holes in the first half month of field planting, uniformly mixing plant ash and bottom soil to return the holes to form soil piles with the height not more than 5cm higher than the ground to field-planted young coffee plants, digging holes in the direction away from the coffee plants by taking the water dripping line of coffee tree crowns as a boundary, stacking the dug surface soil and the dug bottom soil separately, returning the holes after mixing the plant ash and the bottom soil, compacting the surface soil on the holes, and returning the rest surface soil to the heads of the coffee plants. The mode of applying plant ash is limited, so that the root system activity of coffee plants is obviously improved, the full absorption of the plants to nutrients is facilitated, and the utilization rate of the fertilizer is further improved. The success rate of the coffee tree after grafting is improved.

The specific steps of balanced fertilization are as follows: digging an annular shallow ditch at the projection position of the coffee crown in the current year 10-12 months, wherein the ditch depth is 10cm, broadcasting plant ash, the application amount of the plant ash is 0.3-0.5kg, in the next year 10-12 months, collecting a soil sample, drying the soil sample to detect soil nutrients, and combining the nutrients required by the growth of coffee plants in the coffee kingdom according to the mass ratio of N: p₂O₅：K₂O is 1: 0.2-1.5: the required amount of the nitrogenous fertilizer, the phosphate fertilizer and the potash fertilizer is calculated according to the proportion of 0.2-1.5, and the calculation method is that the N content in the nitrogenous fertilizer, the phosphate fertilizer and the potash fertilizer is calculated according to the proportion: p₂O₅：K₂And calculating the content of O, fully and uniformly mixing the three fertilizers to prepare additional fertilizers, ditching two sides of each row of tea trees, applying the phosphorus-potassium fertilizer for the first time in 5 months at intervals of one ditch, applying the phosphorus-potassium fertilizer for the second time in 8 months, and applying the phosphorus-potassium fertilizer for the third time and the fourth time in 3 months and 6 months of the second year respectively, wherein the method is the same as the method for applying the phosphorus-potassium fertilizer for the first time and the second time.

Wherein, as preferred, balanced fertilization still includes the foliage dressing: the boron fertilizer and the zinc fertilizer are prepared into a solution with the concentration of 0.30-0.50% according to the mass ratio of B to Zn to 1 to 3, and the solution is sprayed on the leaf surfaces in 4 months and 8 months every year.

Compared with the traditional fertilizing method, the invention respectively fertilizes at different periods according to the characteristics of the fertilizer required by coffee, improves the effectiveness of the fertilizer, supplements trace elements, provides good nutrition for the growth of coffee plants, provides guarantee for high and stable yield, and can increase the yield by 15-18% on the original basis.

The method combines the surface application of macroelements and the foliage spraying of trace elements, meets the requirements of coffee on different elements, greatly improves the effectiveness of the fertilizer, can obviously improve the yield of coffee, reduces the loss of the fertilizer and saves the production cost.

The management of coffee also includes pruning: trimming all coffee trees, randomly selecting a row, cutting off branches and leaves at the position, 60-70 cm away from the ground, of 2-6 coffee trees at one end to form a bundle of short fir trees, cutting off branches and leaves at 2-3 leaf layers of the adjacent 2-6 coffee tree surfaces in the same row to form a bundle of high fir trees, and so on, wherein the short fir trees and the high fir trees are alternately arranged, the processing modes of the adjacent rows of coffee trees are opposite, namely, the branches and leaves at the 2-3 leaf layers of the adjacent 2-6 tree surfaces at the front end are cut off to form a bundle of high fir trees, then the branches and leaves at the position, 60-70 cm away from the ground, of the adjacent 2-6 trees in the same row are cut off to form a bundle of short fir trees, and so on, the high fir trees and the low fir trees are alternately arranged; and carrying out second trimming in 10 months, wherein the height of the second trimming is increased by 15-20 cm on the trimming opening of the first trimming, and the rest processing modes are the same as the first trimming.