CN114651658A - Creation and identification method of cold-resistant seeds of macadimia nuts - Google Patents
Creation and identification method of cold-resistant seeds of macadimia nuts Download PDFInfo
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Abstract
The invention discloses a method for creating and identifying cold-resistant seeds of macadimia nuts, which comprises the following steps: (1) seed selection; (2) seed soaking treatment; (3) carrying out temperature change treatment; (4) seedling raising; (5) identifying and selecting products; performing mutation breeding on the macadimia nuts by combining temperature change treatment and low-temperature chemical mutation treatment, and performing the seedling culture process under a lower temperature condition, so that the macadimia nut seedlings with good cold resistance can be preferentially screened out through mutation; the seedling process is carried out on a purpose-made artificial seedbed, the purpose-made artificial seedbed not only can provide sufficient nutrients for the macadamia nut seedlings, but also is reasonable in soil structure, is particularly suitable for longitudinal and transverse growth of the roots of the macadamia nut seedlings, and is convenient for calculating the cold resistance of the macadamia nut seedlings according to the ratio of the depth to the width of the roots of the seedlings while the survival rate of the macadamia nut seedlings is improved.
Description
Technical Field
The invention relates to the technical field of agricultural planting, in particular to a method for creating and identifying cold-resistant seeds of macadimia nuts.
Background
The macadamia nut is also named as Kunzea chestnut, macadamia walnut, macadamia nut and Kunzea fruit, and is a tree nut native to Australia. Macadamia nuts belong to evergreen trees and dicotyledonous plants. The tree crown is tall and big, 3-4 leaves are in rotation, and the tree crown is needle-shaped, is leather, is smooth and has thorn-shaped sawteeth on the edge. Raceme axillary, beige flower, round fruit, pericarp, hard endocarp, and beige to light brown kernel. Is suitable for growing in mild, humid and wind-driven areas. Among the numerous dried fruits in the world, macadamia nut is the most economically valuable, enjoying the reputation of "king of dried fruit". Macadamia nut is rich in nutrition (milk white), its epicarp is green, its endocarp is hard and brown, its single fruit weight is 15-16 g, oil content is about 70%, protein is 9%, and contains 17 amino acids including 8 amino acids necessary for human body, and also contains mineral substances and vitamins. The macadamia nut is crisp, tender and delicious, has unique cream fragrance, is an edible fruit with the best quality in the world, has the beauty of 'dried fruit queen' and 'world nut king', and has far better flavor and mouthfeel than cashew nuts. Macadamia nut can be used for making dried fruits, cakes, chocolates, edible oil, cosmetics and the like. In addition, macadamia nut also has high nutritional value and medicinal value.
At present, the cold resistance of the macadimia nuts is low, and in addition, the temperature of all parts of the macadimia nuts sometimes appears in extreme cold weather in recent years, and the cold damage of the macadimia nuts often occurs, so that the macadimia nut seedlings with simple identification method and good cold resistance effect need to be developed urgently.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide an identification method and a creation and identification method of cold-resistant seeds of macadimia nuts with good cold-resistant effect.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows: a method for creating and identifying cold-resistant seeds of macadimia nuts comprises the following steps:
(1) seed selection: selecting mature macadamia nut with shell, which has no plant diseases and insect pests, smooth surface of fruit shell and no damage as seed;
(2) seed soaking treatment: soaking the seeds in a propylene glycol solution of propyl p-hydroxybenzoate, taking out and drying in the air to obtain surface-treated seeds;
(3) temperature change treatment:
3.1) circularly heating and cooling: adjusting the temperature of the periodic heating and cooling device to 8-12 ℃, and putting the seeds subjected to surface treatment into the periodic heating and cooling device to perform cyclic heating and cooling for 3-5 times to obtain temperature-change treated seeds; the technological parameters of the circulating temperature rise and fall are as follows: reducing the temperature of the device to-8 to-12 ℃ under the conditions that the temperature is 8 to 12 ℃ and the temperature reduction rate is 0.8 to 1.2 ℃/min; then under the conditions that the temperature is-8 to-12 ℃, the heating rate is 0.8 to 1.2 ℃/min, the temperature of the device is increased to 8 to 12 ℃, and 1-time circulation heating and cooling can be completed;
3.2) low-temperature treatment: soaking the seeds subjected to temperature change treatment in a low-temperature treatment solution for 4-6 h, preserving at the temperature of-3 to-5 ℃ for 1-2 h, taking out and airing to obtain the seeds subjected to low-temperature treatment; the low-temperature treatment liquid takes sterile water as a solvent, and contains 120-150 g/kg of ethanol, 1.6-1.8 g/kg of nitrosoethyl urea and 0.3-0.5 g/kg of compound sodium nitrophenolate;
(4) seedling culture: placing the seeds subjected to low-temperature treatment into a second black soil layer of the artificial seedbed, and raising the seedlings to a seedling height of 50-60 cm at 15-18 ℃ to obtain the created seedlings; the artificial seedbed is sequentially provided with a first fine sand layer, a loess layer, a first black soil layer, a second fine sand layer, a plant ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top;
(5) and (4) identifying and selecting products: the ratio of the depth to the width of the root system of the created seedling is used as the cold resistance standard.
Further, the ripe macadamia nut with shell in the step (1) is a fresh fruit picked in the current year, and the variety of the ripe macadamia nut with shell is H2.
Further, in the step (2), the seeds are placed into a propylene glycol solution with propyl p-hydroxybenzoate concentration of 10-20 g/kg for seed soaking for 4-6 h, and the seeds are taken out and dried to obtain the surface-treated seeds.
Further, in the step (3), the temperature changing process:
3.1) circularly heating and cooling: adjusting the temperature of the device for periodically heating and cooling to 10-12 ℃, and putting the seeds subjected to surface treatment into the device for periodically heating and cooling for 4-5 times of cyclic heating and cooling to obtain seeds subjected to temperature change treatment; the technological parameters of the circulating temperature rise and fall are as follows: reducing the temperature of the device to-10 to-12 ℃ under the conditions that the temperature is 10 to 12 ℃ and the temperature reduction rate is 1.0 to 1.2 ℃/min; then under the conditions that the temperature is-10 to-12 ℃, the heating rate is 1.0 to 1.2 ℃/min, the temperature of the device is increased to 10 to 12 ℃, and 1-time circulation heating and cooling can be completed;
3.2) low-temperature treatment: soaking the seeds subjected to temperature change treatment in a low-temperature treatment solution for 5-6 h, preserving at the temperature of-4 to-5 ℃ for 1.5-2 h, taking out and airing to obtain the seeds subjected to low-temperature treatment; the low-temperature treatment liquid takes sterile water as a solvent, and contains 130-150 g/kg of ethanol, 1.7-1.8 g/kg of nitrosoethyl urea and 0.4-0.5 g/kg of compound sodium nitrophenolate.
Further, the low-temperature treatment liquid in the step (3) also contains brassinolide with the concentration of 0.25-0.35 g/kg.
Further, in the step (4), the seeds treated at the low temperature are placed in a second black soil layer of the artificial seedbed, and the seedlings are cultivated at the temperature of 16-18 ℃ until the seedling height is 55-60 cm, so that the created seedlings are obtained.
Further, the artificial seedbed in the step (4) is sequentially provided with a first fine sand layer, a yellow soil layer, a first black soil layer, a second fine sand layer, a grass-wood ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top, wherein the first fine sand layer is 20-30 cm in height, the yellow soil layer is 15-20 cm in height, the first black soil layer is 20-30 cm in height, the second fine sand layer is 10-15 cm in height, the grass-wood ash layer is 5-7 cm in height, the decomposed pig manure layer is 8-10 cm in height, and the second black soil layer is 10-15 cm in height.
Further, the artificial seedbed in the step (4) is sequentially provided with a first fine sand layer, a yellow soil layer, a first black soil layer, a second fine sand layer, a grass-wood ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top, wherein the first fine sand layer is 25-30 cm in height, the yellow soil layer is 17-20 cm in height, the first black soil layer is 25-30 cm in height, the second fine sand layer is 12-15 cm in height, the grass-wood ash layer is 6-7 cm in height, the decomposed pig manure layer is 9-10 cm in height, and the second black soil layer is 12-15 cm in height.
Further, the cold tolerance standard in the step (5) is as follows: when the ratio of the root depth to the root width is 1.7-1.9, the cold resistance is strong; when the ratio of the root depth to the root width is 1.5-1.6, the cold resistance is general; when the ratio of the root depth to the root width is 2.0-2.2, the cold resistance is weak.
The invention has the beneficial effects that: the invention relates to a method for creating and identifying cold-resistant seeds of macadimia nuts, which is characterized in that the seeds of the macadimia nuts are created by adopting chemical mutagenesis or ray irradiation mutagenesis and the like at present to carry out mutation breeding, the seeds obtained by the chemical mutagenesis or the ray irradiation mutagenesis have various shapes, but do not necessarily have excellent cold resistance, so that the mutant breeding of the macadimia nuts is carried out by combining temperature-change treatment and low-temperature chemical mutagenesis treatment, and the seedling culture process is carried out under the condition of lower temperature, so that the seedlings of the macadimia nuts with good cold-resistant effect can be preferentially mutagenized and screened; the seedling process is carried out on a purpose-made artificial seedbed, the purpose-made artificial seedbed not only can provide sufficient nutrients for the macadamia nut seedlings, but also is reasonable in soil structure, is particularly suitable for longitudinal and transverse growth of the roots of the macadamia nut seedlings, and is convenient for calculating the cold resistance of the macadamia nut seedlings according to the ratio of the depth to the width of the roots of the seedlings while the survival rate of the macadamia nut seedlings is improved.
According to the method for creating and identifying the cold-resistant germplasm of the macadimia nut, seeds are put into a propylene glycol solution of propyl p-hydroxybenzoate for soaking, so that the sterilization treatment of the macadimia nut seeds can be completed, pathogenic bacteria on the upper surface of the macadimia nut seeds can be killed, and the emergence rate is higher; the seeds are subjected to cyclic heating and cooling, and then the seeds subjected to temperature change are subjected to low-temperature chemical mutagenesis treatment, so that the effect of strengthening the chemical mutagenesis effect can be achieved, the chemical mutagenesis of the macadimia nut seeds is promoted, and the yield of cold-resistant macadimia nut seedlings can be increased; nitrosoethyl urea in the low-temperature treatment liquid is an alkylating agent, the action mechanism is alkylation, hydrogen atoms of other molecules are replaced through alkyl replacement, the action is mainly nucleic acid, DNA (deoxyribonucleic acid) breakage, deletion or repair can be caused, and a powerful cell activator, namely compound sodium nitrophenolate, is added, so that the protoplasm flow of cells can be promoted, the cell activity is improved, the mutation speed of macadamia nut seeds is accelerated, the mutation rate is improved, and the seeds with the cold-resistant property can be obtained.
According to the creating and identifying method of the cold-resistant germplasm of the macadimia nuts, the variety of mature macadimia nuts with shells is H2, other cold-resistant varieties can also be selected as standby seeds, and due to the steps of cyclic temperature rise and fall, low-temperature treatment, low-temperature seedling culture and the like in the operation process, when the cold-resistant variety of the macadimia nut seeds is selected, cold-resistant seedlings are more easily created, and the survival rate of the created seedlings is higher; brassinolide is also added into the low-temperature treatment liquid, and the brassinolide is used as a growth regulator, so that the inherent potential of the macadamia nut seeds can be fully excited, and the effect of improving the survival rate of the mutagenized seeds is achieved.
Detailed Description
The following examples may help one skilled in the art to more fully understand the present invention, but are not intended to limit the invention in any way.
The invention relates to a method for creating and identifying cold-resistant seeds of macadimia nuts, which is characterized in that in order to obtain a better sterilization effect, the volume of propyl p-hydroxybenzoate solution is slightly larger than the volume of seeds, and the dosage of the propyl p-hydroxybenzoate solution can be selected according to actual conditions; other common disinfectant can also be selected for sterilizing the seeds.
The invention relates to a method for creating and identifying cold-resistant seeds of macadimia nuts.
Example 1
A method for creating and identifying cold-resistant seeds of macadimia nuts comprises the following steps:
(1) seed selection: selecting mature macadamia nut with shell which is free from plant diseases and insect pests, smooth in shell surface, free from damage and picked in the same year as seeds; the varieties of mature macadamia nut with shell are H2 and HAES 900 respectively;
(2) seed soaking treatment: soaking the seeds in propylene glycol solution with propyl p-hydroxybenzoate concentration of 10g/kg for 4h, taking out, and air drying to obtain surface-treated seeds; the volume ratio of the seeds to the propyl p-hydroxybenzoate solution is 1: 5;
(3) temperature change treatment:
3.1) circularly heating and cooling: adjusting the temperature of the device for periodically heating and cooling to 8 ℃, and putting the seeds subjected to surface treatment into the device for periodically heating and cooling to perform heating and cooling for 3 times in a circulating manner to obtain seeds subjected to temperature change treatment; the technological parameters of the circulating temperature rise and fall are as follows: reducing the temperature of the device to-8 ℃ under the conditions that the temperature is 8 ℃ and the cooling rate is 0.8 ℃/min; then under the conditions that the temperature is-8 ℃ and the heating rate is 0.8 ℃/min, the temperature of the device is raised to 8 ℃, and 1-time circulation heating and cooling can be completed;
3.2) low-temperature treatment: soaking the seeds subjected to temperature change treatment in the low-temperature treatment solution for 4h, preserving at-3 ℃ for 1h, taking out and drying in the air to obtain low-temperature treated seeds; the low-temperature treatment liquid takes sterile water as a solvent and contains 120g/kg of ethanol, 1.6g/kg of nitrosoethyl urea, 0.3g/kg of compound sodium nitrophenolate and 0.25g/kg of brassinolide;
(4) seedling culture: placing the seeds treated at the low temperature into a second black soil layer of the artificial seedbed, and raising the seedlings to be 50 cm higher than the seedlings at the temperature of 15 ℃ to obtain the created seedlings; the artificial seedbed is sequentially provided with a first fine sand layer, a yellow soil layer, a first black soil layer, a second fine sand layer, a grass and wood ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top, wherein the first fine sand layer is 20 cm in height, the yellow soil layer is 15 cm in height, the first black soil layer is 20 cm in height, the second fine sand layer is 10 cm in height, the grass and wood ash layer is 5 cm in height, the decomposed pig manure layer is 8 cm in height, and the second black soil layer is 10 cm in height;
(5) and (4) identifying and selecting products: taking the ratio of the depth to the width of the root system of the created seedling as the cold resistance standard; the cold resistance criteria were: when the ratio of the root depth to the root width is 1.7-1.9, the cold resistance is strong; when the ratio of the root depth to the root width is 1.5-1.6, the cold resistance is general; when the ratio of the root depth to the root width is 2.0-2.2, the cold resistance is weak.
Example 2
A method for creating and identifying cold-resistant seeds of macadimia nuts comprises the following steps:
(1) seed selection: selecting mature macadamia nut with shell which is free from plant diseases and insect pests, smooth in shell surface, free from damage and picked in the same year as seeds; the varieties of mature macadamia nut with shell are H2 and HAES 900 respectively;
(2) seed soaking treatment: soaking the seeds in a propylene glycol solution with the concentration of the propyl p-hydroxybenzoate being 20g/kg for 6 hours, taking out and airing to obtain seeds with surface treatment; the volume ratio of the seeds to the propyl p-hydroxybenzoate solution is 1: 5;
(3) temperature change treatment:
3.1) circularly heating and cooling: adjusting the temperature of the device for periodically heating and cooling to 12 ℃, and putting the seeds subjected to surface treatment into the device for periodically heating and cooling to perform circulation heating and cooling for 5 times to obtain seeds subjected to temperature change treatment; the technological parameters of the circulating temperature rise and fall are as follows: reducing the temperature of the device to-12 ℃ under the conditions that the temperature is 12 ℃ and the cooling rate is 1.2 ℃/min; then under the conditions that the temperature is-12 ℃ and the heating rate is 1.2 ℃/min, the temperature of the device is raised to 12 ℃, and 1-time circulation heating and cooling can be completed;
3.2) low-temperature treatment: soaking the seeds subjected to temperature change treatment in the low-temperature treatment solution for 6h, preserving at-5 ℃ for 2h, taking out and drying in the air to obtain low-temperature treated seeds; the low-temperature treatment liquid takes sterile water as a solvent, and contains 150g/kg of ethanol, 1.8g/kg of nitrosoethyl urea, 0.5g/kg of compound sodium nitrophenolate and 0.35g/kg of brassinolide;
(4) seedling culture: placing the seeds treated at the low temperature into a second black soil layer of the artificial seedbed, and culturing the seedlings at 18 ℃ until the seedling height is 60 cm, so as to obtain the created seedlings; the artificial seedbed is sequentially provided with a first fine sand layer, a yellow soil layer, a first black soil layer, a second fine sand layer, a grass and wood ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top, wherein the first fine sand layer is 30 cm in height, the yellow soil layer is 20 cm in height, the first black soil layer is 30 cm in height, the second fine sand layer is 15 cm in height, the grass and wood ash layer is 7 cm in height, the decomposed pig manure layer is 10 cm in height, and the second black soil layer is 15 cm in height;
(5) and (4) identifying and selecting products: taking the ratio of the depth to the width of the root system of the created seedling as the cold resistance standard; the cold resistance criteria were: when the ratio of the root depth to the root width is 1.7-1.9, the cold resistance is strong; when the ratio of the root depth to the root width is 1.5-1.6, the cold resistance is general; when the ratio of the root depth to the root width is 2.0-2.2, the cold resistance is weak.
Example 3
A method for creating and identifying cold-resistant seeds of macadimia nuts comprises the following steps:
(1) seed selection: selecting mature macadimia nuts with shells, which are free from plant diseases and insect pests, smooth in shell surface, free from damage and picked in the same year, as seeds; the varieties of mature macadimia nut with shell are respectively H2 and HAES 900;
(2) seed soaking treatment: soaking the seeds in a propylene glycol solution with the propyl p-hydroxybenzoate concentration of 15g/kg for 5h, taking out and airing to obtain surface-treated seeds; the volume ratio of the seeds to the propyl p-hydroxybenzoate solution is 1: 5;
(3) temperature changing treatment:
3.1) circularly heating and cooling: adjusting the temperature of the device for periodically heating and cooling to 10 ℃, and putting the seeds subjected to surface treatment into the device for periodically heating and cooling to perform circulation heating and cooling for 4 times to obtain seeds subjected to temperature change treatment; the technological parameters of the circulating temperature rise and fall are as follows: reducing the temperature of the device to-10 ℃ under the conditions that the temperature is 10 ℃ and the cooling rate is 1.0 ℃/min; then under the conditions that the temperature is minus 10 ℃ and the heating rate is 1.0 ℃/min, the temperature of the device is raised to 10 ℃, and 1-time circulation heating and cooling can be completed;
3.2) low-temperature treatment: soaking the seeds subjected to temperature change treatment in the low-temperature treatment solution for 5h, preserving at-4 ℃ for 1.5h, taking out and drying in the air to obtain low-temperature treated seeds; the low-temperature treatment liquid takes sterile water as a solvent and contains 130g/kg of ethanol, 1.7g/kg of nitrosoethyl urea, 0.4g/kg of compound sodium nitrophenolate and 0.28g/kg of brassinolide;
(4) seedling culture: the seeds treated at low temperature are placed in a second black soil layer of the artificial seedbed, and the seedlings are cultured at the temperature of 16 ℃ until the seedling height is 55 cm, so that the created seedlings are obtained; the artificial seedbed is sequentially provided with a first fine sand layer, a yellow soil layer, a first black soil layer, a second fine sand layer, a grass-wood ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top, wherein the first fine sand layer is 25 cm in height, the yellow soil layer is 17 cm in height, the first black soil layer is 25 cm in height, the second fine sand layer is 12 cm in height, the grass-wood ash layer is 6 cm in height, the decomposed pig manure layer is 9 cm in height, and the second black soil layer is 12 cm in height;
(5) and (4) identifying and selecting products: taking the ratio of the depth to the width of the root system of the created seedling as the cold resistance standard; the cold resistance criteria were: when the ratio of the root depth to the root width is 1.7-1.9, the cold resistance is strong; when the ratio of the root depth to the root width is 1.5-1.6, the cold resistance is general; when the ratio of the root depth to the root width is 2.0-2.2, the cold resistance is weak.
Example 4
A method for creating and identifying cold-resistant seeds of macadimia nuts comprises the following steps:
(1) seed selection: selecting mature macadamia nut with shell which is free from plant diseases and insect pests, smooth in shell surface, free from damage and picked in the same year as seeds; the varieties of mature macadimia nut with shell are respectively H2 and HAES 900;
(2) seed soaking treatment: soaking the seeds in a propylene glycol solution with the propyl p-hydroxybenzoate concentration of 15g/kg for 5h, taking out and airing to obtain surface-treated seeds; the volume ratio of the seeds to the propyl p-hydroxybenzoate solution is 1: 5;
(3) temperature change treatment:
3.1) circularly heating and cooling: adjusting the temperature of the device for periodically heating and cooling to 10 ℃, and putting the seeds subjected to surface treatment into the device for periodically heating and cooling to perform circulation heating and cooling for 4 times to obtain seeds subjected to temperature change treatment; the technological parameters of the circulating temperature rise and fall are as follows: reducing the temperature of the device to-10 ℃ under the conditions that the temperature is 10 ℃ and the cooling rate is 1.0 ℃/min; then under the conditions that the temperature is minus 10 ℃ and the heating rate is 1.0 ℃/min, the temperature of the device is raised to 10 ℃, and 1-time circulation heating and cooling can be completed;
3.2) low-temperature treatment: soaking the seeds subjected to temperature change treatment in the low-temperature treatment solution for 5h, preserving at-4 ℃ for 1.5h, taking out and drying in the air to obtain low-temperature treated seeds; the low-temperature treatment liquid takes sterile water as a solvent, and contains 130g/kg of ethanol, 1.7g/kg of nitrosoethylurea and 0.4g/kg of compound sodium nitrophenolate;
(4) seedling culture: the seeds treated at low temperature are placed in a second black soil layer of the artificial seedbed, and the seedlings are cultured at the temperature of 16 ℃ until the seedling height is 55 cm, so that the created seedlings are obtained; the artificial seedbed is sequentially provided with a first fine sand layer, a yellow soil layer, a first black soil layer, a second fine sand layer, a grass-wood ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top, wherein the first fine sand layer is 25 cm in height, the yellow soil layer is 17 cm in height, the first black soil layer is 25 cm in height, the second fine sand layer is 12 cm in height, the grass-wood ash layer is 6 cm in height, the decomposed pig manure layer is 9 cm in height, and the second black soil layer is 12 cm in height;
(5) and (4) identifying and selecting products: taking the ratio of the depth to the width of the root system of the created seedling as the cold resistance standard; the cold resistance criteria were: when the ratio of the root depth to the root width is 1.7-1.9, the cold resistance is strong; when the ratio of the root depth to the root width is 1.5-1.6, the cold resistance is general; when the ratio of the root depth to the root width is 2.0-2.2, the cold resistance is weak.
Comparative example 1
A method for creating and identifying cold-resistant seeds of macadimia nuts comprises the following steps:
(1) seed selection: selecting mature macadimia nuts with shells, which are free from plant diseases and insect pests, smooth in shell surface, free from damage and picked in the same year, as seeds; the varieties of mature macadamia nut with shell are H2 and HAES 900 respectively;
(2) seed soaking treatment: soaking the seeds in a propylene glycol solution with the propyl p-hydroxybenzoate concentration of 15g/kg for 5h, taking out and airing to obtain surface-treated seeds; the volume ratio of the seeds to the propyl p-hydroxybenzoate solution is 1: 5;
(3) temperature change treatment:
3.1) circularly heating and cooling: adjusting the temperature of the device for periodically heating and cooling to 10 ℃, and putting the seeds subjected to surface treatment into the device for periodically heating and cooling to perform circulation heating and cooling for 4 times to obtain seeds subjected to temperature change treatment; the technological parameters of the circulating temperature rise and fall are as follows: reducing the temperature of the device to-10 ℃ under the conditions that the temperature is 10 ℃ and the cooling rate is 1.0 ℃/min; then under the conditions that the temperature is minus 10 ℃ and the heating rate is 1.0 ℃/min, the temperature of the device is raised to 10 ℃, and 1-time circulation heating and cooling can be completed;
3.2) low-temperature treatment: soaking the seeds subjected to temperature change treatment in the low-temperature treatment solution for 5h, preserving at-4 ℃ for 1.5h, taking out and drying in the air to obtain low-temperature treated seeds; the low-temperature treatment liquid takes sterile water as a solvent and contains 130g/kg of ethanol, 1.7g/kg of nitrosoethyl urea, 0.4g/kg of compound sodium nitrophenolate and 0.28g/kg of brassinolide;
(4) seedling culture: placing the seeds treated at low temperature into a traditional indoor seedbed, and raising the seedlings to 55 cm higher than the seedlings at 16 ℃ to obtain the created seedlings;
(5) and (4) identifying and selecting products: taking the ratio of the depth to the width of the root system of the created seedling as the cold resistance standard; the cold resistance standard is as follows: when the ratio of the root depth to the root width is 1.7-1.9, the cold resistance is strong; when the ratio of the root depth to the root width is 1.5-1.6, the cold resistance is general; when the ratio of the root depth to the root width is 2.0-2.2, the cold resistance is weak.
Comparative example 2
A method for creating and identifying cold-resistant seeds of macadimia nuts comprises the following steps:
(1) seed selection: selecting mature macadamia nut with shell which is free from plant diseases and insect pests, smooth in shell surface, free from damage and picked in the same year as seeds; the varieties of mature macadamia nut with shell are H2 and HAES 900 respectively;
(2) seed soaking treatment: putting the seeds into a propylene glycol solution with propyl p-hydroxybenzoate concentration of 15g/kg for soaking the seeds for 5 hours, taking out and airing to obtain seeds with surface treatment; the volume ratio of the seeds to the propyl p-hydroxybenzoate solution is 1: 5;
(3) low-temperature treatment:
low-temperature treatment: soaking the surface-treated seeds in the low-temperature treatment solution for 5h, preserving at-4 ℃ for 1.5h, taking out and airing to obtain low-temperature-treated seeds; the low-temperature treatment liquid takes sterile water as a solvent and contains 130g/kg of ethanol, 1.7g/kg of nitrosoethyl urea, 0.4g/kg of compound sodium nitrophenolate and 0.28g/kg of brassinolide;
(4) seedling culture: the seeds treated at low temperature are placed in a second black soil layer of the artificial seedbed, and the seedlings are cultured at the temperature of 16 ℃ until the seedling height is 55 cm, so that the created seedlings are obtained; the artificial seedbed is sequentially provided with a first fine sand layer, a yellow soil layer, a first black soil layer, a second fine sand layer, a grass-wood ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top, wherein the first fine sand layer is 25 cm in height, the yellow soil layer is 17 cm in height, the first black soil layer is 25 cm in height, the second fine sand layer is 12 cm in height, the grass-wood ash layer is 6 cm in height, the decomposed pig manure layer is 9 cm in height, and the second black soil layer is 12 cm in height;
(5) and (3) identifying and selecting products: taking the ratio of the depth to the width of the root system of the created seedling as the cold resistance standard; the cold resistance criteria were: when the ratio of the root depth to the root width is 1.7-1.9, the cold resistance is strong; when the ratio of the root depth to the root width is 1.5-1.6, the cold resistance is general; when the ratio of the root depth to the root width is 2.0-2.2, the cold resistance is weak.
Comparative example 3
A method for creating and identifying cold-resistant seeds of macadimia nuts comprises the following steps:
(1) seed selection: selecting mature macadamia nut with shell which is free from plant diseases and insect pests, smooth in shell surface, free from damage and picked in the same year as seeds; the varieties of mature macadamia nut with shell are H2 and HAES 900 respectively;
(2) seed soaking treatment: soaking the seeds in a propylene glycol solution with the propyl p-hydroxybenzoate concentration of 15g/kg for 5h, taking out and airing to obtain surface-treated seeds; the volume ratio of the seeds to the propyl p-hydroxybenzoate solution is 1: 5;
(3) temperature change treatment:
3.1) circularly heating and cooling: adjusting the temperature of the device for periodically heating and cooling to 10 ℃, and putting the seeds subjected to surface treatment into the device for periodically heating and cooling to perform circulation heating and cooling for 4 times to obtain seeds subjected to temperature change treatment; the technological parameters of the circulating temperature rise and fall are as follows: reducing the temperature of the device to-10 ℃ under the conditions that the temperature is 10 ℃ and the cooling rate is 1.0 ℃/min; then under the conditions that the temperature is minus 10 ℃ and the heating rate is 1.0 ℃/min, the temperature of the device is raised to 10 ℃, and 1-time circulation heating and cooling can be completed;
3.2) low-temperature treatment: soaking the seeds subjected to temperature change treatment in the low-temperature treatment solution for 5h, preserving at-4 ℃ for 1.5h, taking out and drying in the air to obtain low-temperature treated seeds; the low-temperature treatment liquid takes sterile water as a solvent, and contains 130g/kg of ethanol, 1.7g/kg of nitrosoethyl urea and 0.28g/kg of brassinolide;
(4) seedling culture: placing the seeds treated at the low temperature into a second black soil layer of the artificial seedbed, and culturing the seedlings at the temperature of 16 ℃ until the seedling height is 55 cm, so as to obtain the created seedlings; the artificial seedbed is sequentially provided with a first fine sand layer, a yellow soil layer, a first black soil layer, a second fine sand layer, a grass-wood ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top, wherein the first fine sand layer is 25 cm in height, the yellow soil layer is 17 cm in height, the first black soil layer is 25 cm in height, the second fine sand layer is 12 cm in height, the grass-wood ash layer is 6 cm in height, the decomposed pig manure layer is 9 cm in height, and the second black soil layer is 12 cm in height;
(5) and (4) identifying and selecting products: taking the ratio of the depth to the width of the root system of the created seedling as the cold resistance standard; the cold resistance criteria were: when the ratio of the root depth to the root width is 1.7-1.9, the cold resistance is strong; when the ratio of the root depth to the root width is 1.5-1.6, the cold resistance is general; when the ratio of the root depth to the root width is 2.0-2.2, the cold resistance is weak.
The experimental results of the creation and identification of cold-resistant seeds of macadamia nuts in examples 1 to 4 and comparative examples 1 to 3 were counted:
the method for calculating the number of the abnormal seedlings comprises the following steps: observing the form of the seedlings, observing special characters of the created seedlings, specifically dwarf plants, leaf deformity, yellowing of leaves, poor cold resistance and the like, counting data of the special characters, and calculating the number of the distorted seedlings;
the detection method of cold resistance comprises the following steps: a conductivity method, which is used for identifying whether the created seedlings have cold resistance; certainly, the created seedlings can also be placed in a low-temperature environment for cultivation, and the cold resistance of the created seedlings is judged by observing the occurrence condition of low-temperature cold damage;
the following table 1 shows a statistical table of the results of the germination rates, the survival rates of created seedlings, the distortion numbers, and the cold-resistant seedlings numbers of the seeds in examples 1 to 4 and comparative examples 1 to 3;
TABLE 1
According to the experimental results, the method for creating and identifying the cold-resistant seeds of the macadimia nuts is characterized in that the macadimia nuts are subjected to mutation breeding by combining temperature change treatment and low-temperature chemical mutation treatment, and the seedling culture process is carried out under the condition of lower temperature, so that the macadimia nut seedlings with good cold-resistant effect can be screened out through preferential mutation; the seeds are subjected to cyclic heating and cooling, and then the seeds subjected to temperature change are subjected to low-temperature chemical mutagenesis treatment, so that the effect of strengthening the chemical mutagenesis effect can be achieved, the chemical mutagenesis of the macadimia nut seeds is promoted, and the yield of cold-resistant macadimia nut seedlings can be increased; nitrosoethyl urea in the low-temperature treatment liquid is an alkylating agent, the action mechanism is alkylation, hydrogen atoms of other molecules are replaced through alkyl replacement, the action is mainly nucleic acid, DNA (deoxyribonucleic acid) breakage, deletion or repair can be caused, and a strong cell activator, namely compound sodium nitrophenolate, is added, so that the protoplasm flow of cells can be promoted, the cell activity is improved, the mutation speed of macadamia nut seeds is accelerated, the mutation rate is improved, and the seeds with the cold-resistant property can be obtained; the cold resistance of H2 is obviously superior to HAES 900, and the data in Table 1 show that when cold-resistant variety macadamia nut seeds are selected, cold-resistant seedlings are easier to create, and the survival rate of the created seedlings is higher; brassinolide is also added into the low-temperature treatment liquid, and the brassinolide is used as a growth regulator, so that the inherent potential of the macadamia nut seeds can be fully excited, and the effect of improving the survival rate of the mutagenized seeds is achieved.
In the embodiments 1 to 4 and the comparative examples 1 to 3, created seedlings with cold resistance are created, whether the created seedlings have cold resistance is identified by a conductivity method, in order to obtain a simple method for identifying the cold resistance of the created seedlings, the method also counts the characters of the created seedlings with cold resistance, particularly the leaf characters and the longitudinal and transverse growth conditions of root systems of the created seedlings, finds out dominant characters capable of quickly screening the created seedlings with cold resistance, and is convenient for identifying cold-resistant seeds of macadamia nuts; leaves of partial cold-resistant created seedlings are long and narrow, but leaves of partial cold-resistant created seedlings have no obvious change, so that whether created seedlings have cold resistance or not is difficult to judge through whether leaves of created seedlings are long and narrow; through continuous research, the method of conductivity method, low-temperature cold injury cultivation method for creating seedlings and the like are mutually proved, and when the ratio of the root depth to the root width is 1.7-1.9, the cold resistance is high; when the ratio of the root depth to the root width is 1.5-1.6, the cold resistance is general; when the ratio of the root depth to the root width is 2.0-2.2, the cold resistance is weak; it should be noted that the root depth here refers to the depth of the root system penetrating into the soil, and the root width refers to the span of the root system in the width direction, that is, the transverse width of the root system.
According to the statistical results of the characters of the cold-resistant created seedlings created in the examples 1 to 4 and the comparative examples 1 to 3, it can be known from the longitudinal and transverse growth conditions of the root systems of the created seedlings in the comparative examples 1 to 4 and the comparative example 1 that the longitudinal depth and the transverse width of the root systems of the created seedlings in the comparative example 1 are lower than those obtained by the statistics of the examples 1 to 4, and therefore, the seedling raising process is performed on a special artificial seedbed, the special artificial seedbed can provide sufficient nutrients for the macadamia seedling, the soil structure of the special artificial seedbed is reasonable, the special artificial seedbed is particularly suitable for the longitudinal and transverse growth of the root systems of the macadamia seedling, the survival rate of the macadamia seedling is improved, and meanwhile, the cold resistance of the macadamia seedling can be conveniently calculated according to the ratio of the depth to the width of the root systems of the seedling.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (9)
1. A method for creating and identifying cold-resistant seeds of macadimia nuts is characterized by comprising the following steps:
(1) seed selection: selecting mature macadamia nut with shell, which has no plant diseases and insect pests, smooth surface of fruit shell and no damage as seed;
(2) seed soaking treatment: soaking the seeds in a propylene glycol solution of propyl p-hydroxybenzoate, taking out and drying in the air to obtain surface-treated seeds;
(3) temperature change treatment:
3.1) circularly heating and cooling: adjusting the temperature of the device for periodically heating and cooling to 8-12 ℃, and putting the seeds subjected to surface treatment into the device for periodically heating and cooling for 3-5 times to circularly heat and cool so as to obtain seeds subjected to temperature change treatment; the technological parameters of the circulating temperature rise and fall are as follows: reducing the temperature of the device to-8 to-12 ℃ under the conditions that the temperature is 8 to 12 ℃ and the temperature reduction rate is 0.8 to 1.2 ℃/min; then under the conditions that the temperature is-8 to-12 ℃, the heating rate is 0.8 to 1.2 ℃/min, the temperature of the device is increased to 8 to 12 ℃, and 1-time circulation heating and cooling can be completed;
3.2) low-temperature treatment: soaking the seeds subjected to temperature change treatment in a low-temperature treatment solution for 4-6 h, preserving at the temperature of-3 to-5 ℃ for 1-2 h, taking out and airing to obtain the seeds subjected to low-temperature treatment; the low-temperature treatment liquid takes sterile water as a solvent, and contains 120-150 g/kg of ethanol, 1.6-1.8 g/kg of nitrosoethyl urea and 0.3-0.5 g/kg of compound sodium nitrophenolate;
(4) seedling culture: placing the seeds subjected to low-temperature treatment into a second black soil layer of the artificial seedbed, and raising the seedlings to a seedling height of 50-60 cm at 15-18 ℃ to obtain the created seedlings; the artificial seedbed is sequentially provided with a first fine sand layer, a loess layer, a first black soil layer, a second fine sand layer, a plant ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top;
(5) and (4) identifying and selecting products: the ratio of the depth to the width of the root system of the created seedling is used as the cold resistance standard.
2. The method of claim 1, wherein the mature shelled macadamia nut in step (1) is fresh fruit harvested in the same year, and the variety of mature shelled macadamia nut is H2.
3. The method for creating and identifying cold-resistant germplasm of macadimia nuts, according to claim 1, wherein in the step (2), the seeds are put into a propylene glycol solution with propyl p-hydroxybenzoate concentration of 10-20 g/kg for 4-6 h, and then are taken out and dried to obtain the surface-treated seeds.
4. The method of claim 1, wherein the annealing treatment in step (3) comprises:
3.1) circularly heating and cooling: adjusting the temperature of the device for periodically heating and cooling to 10-12 ℃, and putting the seeds subjected to surface treatment into the device for periodically heating and cooling for 4-5 times of cyclic heating and cooling to obtain seeds subjected to temperature change treatment; the technological parameters of the circulating temperature rise and fall are as follows: reducing the temperature of the device to-10 to-12 ℃ under the conditions that the temperature is 10 to 12 ℃ and the temperature reduction rate is 1.0 to 1.2 ℃/min; then under the conditions that the temperature is-10 to-12 ℃, the heating rate is 1.0 to 1.2 ℃/min, the temperature of the device is increased to 10 to 12 ℃, and 1-time circulation heating and cooling can be completed;
3.2) low-temperature treatment: soaking the seeds subjected to temperature change treatment in a low-temperature treatment solution for 5-6 h, preserving at the temperature of-4 to-5 ℃ for 1.5-2 h, taking out and airing to obtain the seeds subjected to low-temperature treatment; the low-temperature treatment liquid takes sterile water as a solvent, and contains 130-150 g/kg of ethanol, 1.7-1.8 g/kg of nitrosoethyl urea and 0.4-0.5 g/kg of compound sodium nitrophenolate.
5. The method for creating and identifying cold-resistant germplasm of macadimia nuts as claimed in claim 1, wherein the low-temperature treatment liquid in the step (3) further comprises brassinolide at a concentration of 0.25-0.35 g/kg.
6. The method for creating and identifying cold-resistant germplasm of macadimia nuts according to claim 1, wherein the seeds subjected to low-temperature treatment in the step (4) are placed in a second black soil layer of an artificial seedbed, and are cultured at the temperature of 16-18 ℃ until the seedling height is 55-60 cm, so that created seedlings are obtained.
7. The method for creating and identifying the cold-resistant germplasm of the macadimia nuts as claimed in claim 1, wherein the artificial seedbed in the step (4) is sequentially provided with a first fine sand layer, a loess layer, a first black soil layer, a second fine sand layer, a grass-wood ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top, wherein the first fine sand layer is 20-30 cm in height, the loess layer is 15-20 cm in height, the first black soil layer is 20-30 cm in height, the second fine sand layer is 10-15 cm in height, the grass-wood ash layer is 5-7 cm in height, the decomposed pig manure layer is 8-10 cm in height, and the second black soil layer is 10-15 cm in height.
8. The method for creating and identifying the cold-resistant germplasm of the macadimia nuts as claimed in claim 7, wherein the artificial seedbed in the step (4) is sequentially provided with a first fine sand layer, a loess layer, a first black soil layer, a second fine sand layer, a grass-wood ash layer, a decomposed pig manure layer and a second black soil layer from bottom to top, wherein the first fine sand layer is 25-30 cm in height, the loess layer is 17-20 cm in height, the first black soil layer is 25-30 cm in height, the second fine sand layer is 12-15 cm in height, the grass-wood ash layer is 6-7 cm in height, the decomposed pig manure layer is 9-10 cm in height, and the second black soil layer is 12-15 cm in height.
9. The method of claim 1, wherein the cold tolerance criteria in step (5) are: when the ratio of the root depth to the root width is 1.7-1.9, the cold resistance is strong; when the ratio of the root depth to the root width is 1.5-1.6, the cold resistance is general; when the ratio of the root depth to the root width is 2.0-2.2, the cold resistance is weak.
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