CN103609450A - Method for accelerating rooting of masson pine tissue culture secondary sprout - Google Patents

Method for accelerating rooting of masson pine tissue culture secondary sprout Download PDF

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CN103609450A
CN103609450A CN201310617394.0A CN201310617394A CN103609450A CN 103609450 A CN103609450 A CN 103609450A CN 201310617394 A CN201310617394 A CN 201310617394A CN 103609450 A CN103609450 A CN 103609450A
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bud
subculture
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root
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姚瑞玲
吴幼媚
蔡玲
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Guangxi Zhuang Autonomous Region Forestry Research Institute
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Guangxi Zhuang Autonomous Region Forestry Research Institute
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Abstract

The invention discloses a method for accelerating the rooting of a masson pine tissue culture secondary sprout. The method comprises the following steps of independently intercepting the strongly-growing semi-lignified masson pine tissue culture secondary sprout with the height of 2-3cm; adopting different culture mediums and a two-step rooting method: firstly inoculating the sprout into a half improved MS culture medium IV which contains naphthylacetic acid, indolebutyric acid and saccharose, inducing adventitious root primordia 21-28d, and then transferring the sprout to a half improved MS culture medium V which contains active carbon or ascorbic acid and the saccharose without auxins to accelerate the expression and elongation of an adventitious root. According to the method, a rooted regenerated plant is transplanted into a peat soil, vermiculite and perlite mixed matrix to be cultured, so that the rooting condition of the secondary sprout is greatly improved; the rooting ratio of the masson pine secondary sprout achieves more than 82.5%, the highest rooting ratio achieves 90.6%, the root system quality is high, the transplantation survival rate achieves more than 85.1%, and the highest survival rate achieves 93.0%; good economic benefit and social benefit are achieved.

Description

A kind of promotion masson pine group training subculture blastogenesis method for root
Technical field
The invention belongs to plant propagation technical field, relate to tissue cultivating and seedling technology, especially a kind of promotion masson pine group training subculture blastogenesis method for root.
Background technology
Masson pine (Pinusmassonianan) originates in China, is that one of important species ， China distribution of the important green barren hill of south China, paper making raw material woods and resin tapping woods, natural land woods is very wide.The seeds high as a kind of comprehensive utilization value, promotion potential is large, masson pine not only can be used to produce three plates, papermaking and chemical fibre industry manufacture, also can be used to produce rosin, the turpentine wet goods raw material of industry.In recent years, along with the intensification of contradictions such as peter out of the non-renewable resources such as the growing life requirement of people and timber resource shortage and oil, ecological, economic and social sustainable development is more and more subject to social concerns.The problems such as, resource security peaceful based on China's ecology and social safety, masson pine, as one of the main industrial cut stock seeds of China and Tree Species as Bio-energy, is greatly developed Masson Pine Plantation necessary.
China, since " six or five " National Key Research Programs, has obtained compared with quantum jump at aspects such as masson pine fine-variety breeding and Fast-growing Cultivation of Cinnamomuns.At present, masson pine breeding mainly be take the sexual propagation of seed orchard and seed production stand as main.Yet sexual propagation exists the problem of differentiation, can not improve to greatest extent genetic improvement gain, and vegetative propagation can overcome the above-mentioned deficiency of sexual propagation.But the current still difficulty of traditional vegetative propagation technique of masson pine (cuttage and grafting) meets the demand of afforestation, restricted the process of Fine Variety Breeding of Pinus massoniana.Modern forestry is particular about fast growing, high-quality and efficient, and the use of plant tissue culture technology is the effective way that realizes masson pine choiceness nursery stock fast breeding.
Horse hair Pinus tissue is cultivated the comparatively seeds of difficulty that take root.Numerous scholar's research showed in the past, take root and conventionally can obtain comparatively desirable rooting rate, and after subculture, its simple bud rooting rate is obviously on the low side with the masson pine simple bud of first culture.As take masson pine mature seed as explant source, and Wu little Qin etc. (2009) and season Kong Shu etc. (2010) are directly taken root simple bud after the first culture through the induction of clump bud and elongation, and its rooting rate is respectively 85% and 92.5%; And Xue Ying (2006) is taken root getting simple bud after subculture is cultivated, its rooting rate is only 26.7%.Realize the seedling industrialized production of masson pine group training, must break through subculture blastogenesis root difficulty bottleneck factor.
Summary of the invention
Object of the present invention, mainly for the low present situation of masson pine group training subculture bud rooting rate, for realizing the large-scale production of masson pine group training seedling, provides a kind of promotion masson pine group training subculture blastogenesis method for root.
The present invention is achieved in that
Promote a masson pine group training subculture blastogenesis method for root, comprise two step rooting methods of clump bud induction, the elongation of clump bud, subculture cultivation, root induction, adventitious root extension, test-tube seedling transplanting, obtain masson pine group training subculture blastogenesis offspring, its operating procedure is as follows:
(1) clump bud induction: by after masson pine seed sterilization, carry out seed germination under aseptic condition, then the long hypocotylar germ free apical bud of intercepting band 0.5cm is explant, carries out clump bud induction in inducing culture I;
(2) clump bud extends: the Multiple Buds inducing is extended in elongation medium II to cultivation;
(3) subculture is cultivated: the single terminal bud that cuts elongation bud is inoculated in shoot proliferation medium III and is bred, obtain subculture bud I; Subculture bud I repeating step (2) is extended, singlely cut the subculture bud I of elongation and be divided into terminal bud and band lateral bud bud section, then in step (3) shoot proliferation medium, continue propagation and obtain subculture bud II, by subculture bud II repeating step (2) and step (3) again, obtain new proliferation and subculture bud; Repetitive cycling step (2) and (3), until obtain a large amount of enough culture of rootage subculture buds;
(4) root induction: the single subculture bud that cuts robust growth, semi-lignified, high 2-3cm, be seeded in the 1/2 modified MS medium IV that contains methyl α-naphthyl acetate (NAA), indolebutyric acid (IBA), sucrose, adventitious root primordia 21-28d is processed in induction;
(5) adventitious root extension: will there is the visible adventive root explant of naked eyes and proceed in the 1/2 modified MS medium V that contains active carbon (AC) or ascorbic acid (Vc), sucrose, and promote expression and the elongation of adventive root;
(6) test-tube seedling transplanting: the Transplantation of Regenerated Plantlets of having taken root is cultivated in peat soil, vermiculite and perlite (1: 1: 1) mixed-matrix to water spray moisturizing.
Above-described its material content of medium I is: 6-BA 2.5-4.0mgL -1, NAA 0.05-0.1mgL -1, sucrose 30000mgL -1, acid hydrolyzed casein 500mgL -1and modified MS medium.
Above-described its material content of medium II is: NAA 0.25-0.4mgL -1or active carbon 3000-5000mgL -1, sucrose 30000mgL -1and modified MS medium.
Above-described its material content of medium III is: 6-BA 1.0-3.0mgL -1, KT 0-1.0mgL -1, NAA 0-0.3mgL -1, sucrose 30000mgL -1and modified MS medium.
Above-described its material content of medium IV is: NAA 0.05-0.1mgL -1, IBA 1.0-2.5mgL -1, sucrose 1.0-2.0% and 1/2 modified MS medium.
Above-described its material content of medium V is: AC 3000-5000mgL -1or Vc 10-30mgL -1, sucrose 2.0% and 1/2 modified MS medium.
Basic composition is of above-described modified MS medium: KNO 31650mgL -1; NH 4nO 3600mgL -1; CaCl 22H 2o260mgL -1; MgSO 47H 2o 370mgL -1; Ca (NO 3) 24H 2o 400mgL -1; KH 2pO 4170mgL -1; MnSO 4h 2o22.3mgL -1; ZnSO 47H 2o8.6mgL -1; CuSO 45H 2o0.025mgL -1; H 3bO 36.2mgL -1; Na 2moO 42H 2o0.025mgL -1; KI 0.83mgL -1; CoCl 26H 2o0.025mgL -1; Cobastab 11.0mgL -1; Cobastab 60.5mgL -1; Nicotinic acid 0.5mgL -1; Glycine 2.0mgL -1; Inositol 200mgL -1.
Above-described cultivation controlled condition is: cultivation temperature 22-26 ℃ on daytime, evening 16-20 ℃, illumination 1500-2000lx, illumination every day 16h.
The present invention has the following advantages with respect to existing technology:
Promotion masson pine group training subculture blastogenesis method for root of the present invention, choose the masson pine group training subculture bud that robust growth, semi-lignified and 2-3cm are high, adopt different medium with two step rooting methods, 21-28d days is processed in first induction, then be forwarded to and there is no growth hormone, but in the medium that contains active carbon or ascorbic acid, greatly promote and improved subculture blastogenesis root situation.Adopt this rooting method, masson pine subculture bud rooting rate reaches more than 82.5%, and the highest rooting rate reaches 90.6%, and root system quality is higher, and transplanting survival rate reaches more than 85.1%, and high-servival rate reaches 93.0%, has good economic benefit and social benefit.
Embodiment
The following examples can make the present invention of those skilled in the art's comprehend, but do not limit the present invention in any way.
embodiment 1
1, experiment material
Take masson pine then ripe cone seed germination germ free apical bud be explant, cone picks up from Ningming of Guangxi portiatree provenance construction masson pine seed production stand the good masson pine family of high yield fat Ningming No. 3.
2, experimental technique
(1) clump bud induction
Aseptic explant is inoculated into improvement MS+6-BA 3.0mgL -1+ NAA 0.1mgL -1+ sucrose 30000mgL -1+ agar 3500mgL -1medium on induce cultivation.Cultivate after 26d, average clump bud induction rate reaches 99.1%.
Basic composition is of described modified MS medium: KNO 31650mgL -1; NH 4nO 3600mgL -1; CaCl 22H 2o260mgL -1; MgSO 47H 2o370mgL -1; Ca (NO 3) 24H 2o400mgL -1; KH 2pO 4170mgL -1; MnSO 4h 2o22.3mgL -1; ZnSO 47H 2o8.6mgL -1; CuSO 45H 2o0.025mgL -1; H 3bO 36.2mgL -1; Na 2moO 42H 2o0.025mgL -1; KI 0.83mgL -1; CoCl 26H 2o0.025mgL -1; Cobastab 11.0mgL -1; Cobastab 60.5mgL -1; Nicotinic acid 0.5mgL -1; Glycine 2.0mgL -1; Inositol 200mgL -1.
(2) clump bud extends
The budlet that grows thickly inducing is transferred to improvement MS+NAA 0.25mgL -1+ sucrose 30000mgL -1+ agar 3500mgL -1in elongation medium, to promote the elongation growth of culture.Extend and cultivate 27d, the high 3-5cm of clump bud.
(3) subculture bud is cultivated
The simple bud of plant height 2-3cm in Multiple Buds is cut, be inoculated into modified MS medium+6-BA 2.0mgL -1+ KT 1.0mgL -1+ NAA 0.2mgL -1+ sucrose 30000mgL -1+ agar 3500mgL -1on carry out subculture bud propagation 23d.
Shoot proliferation bud is put in step (2) elongation medium, to promote shoot proliferation bud to extend.Extend and cultivate 28d, the high 3-5cm of shoot proliferation bud.Elongation subculture bud is divided into after the long terminal bud of 2cm and the long band of 1cm lateral bud bud section, and repeating step (3) shoot proliferation and step (2) clump bud extend.Subculture is cultivated 6 times, and average growth coefficient 6.4 obtains large quantities of for taking root subculture bud.
(4) root induction
The single subculture bud that cuts robust growth, semi-lignified, high 2-3cm, is inoculated into 1/2 modified MS medium+NAA 0.1mgL -1+ IBA 2.5mgL -1the former base of inducing adventitious root in+sucrose 1.0% medium.After induction 21d, root induction rate reaches 90.6%.
(5) adventitious root extension
To obviously there is the visible adventive root explant of naked eyes and proceed to 1/2 modified MS medium+AC 5000mgL -1in+sucrose 2.0% medium, promote adventive root express and extend, after 28d, add up adventive root incidence, record mean elements and observe root system quality (between rhizome callus a situation arises).Concrete data are in Table 1.
(6) test-tube seedling transplanting
Regeneration plant root agar is cleaned, transplanted in peat soil, vermiculite and perlite weight ratio and be: in the mixed-matrix of 1: 1: 1, cultivate, water spray moisturizing, added up transplanting survival rate after one month.Concrete data are in Table 1.
The condition of culture temperature of said process: 26 ℃ of daytimes, 20 ℃ of evenings, illumination 2000lx, illumination every day 16h.
embodiment 2
1, experiment material
Take masson pine then ripe cone seed germination germ free apical bud be explant, cone picks up from Ningming of Guangxi portiatree provenance construction masson pine seed production stand the good masson pine family of high yield fat Ningming No. 4.
2, experimental technique
(1) clump bud induction
Aseptic explant is inoculated into improvement MS+6-BA 4.0mgL -1+ NAA 0.1mgL -1+ sucrose 30000mgL -1+ agar 3500mgL -1medium on induce cultivation.Cultivate after 26d, average clump bud induction rate reaches 98.4%.
The basic composition of described modified MS medium is with embodiment 1.
(2) clump bud extends
The budlet that grows thickly inducing is transferred to improvement MS+NAA 0.3mgL -1+ sucrose 30000mgL -1+ agar 3500mgL -1in elongation medium, to promote the elongation growth of culture.Extend and cultivate 28d, the high 3-5cm of clump bud.
(3) subculture bud is cultivated
The simple bud of plant height 2-3cm in Multiple Buds is cut, be inoculated into modified MS medium+6-BA 2.5mgL -1+ KT 0.5mgL -1+ sucrose 30000mgL -1+ agar 3500mgL -1on carry out subculture bud propagation 26d.
Shoot proliferation bud is put in step (2) elongation medium, to promote shoot proliferation bud to extend.Extend and cultivate 25d, the high 3-5cm of shoot proliferation bud.Elongation subculture bud is divided into after the long terminal bud of 2cm and the long band of 1cm lateral bud bud section, and repeating step (3) shoot proliferation and step (2) clump bud extend.Subculture is cultivated 5 times, average growth coefficient 6.0.
(4) root induction
The single subculture bud that cuts robust growth, semi-lignified, high 2-3cm, is inoculated into 1/2 modified MS medium+NAA 0.1mgL -1+ IBA 2.5mgL -1the former base of inducing adventitious root in+sucrose 1.0% medium.After induction 22d, root induction rate reaches 89.8%.
(5) adventitious root extension
To obviously there is the visible adventive root explant of naked eyes and proceed to 1/2 modified MS medium+Vc 10mgL -1in+sucrose 2.0% medium, promote adventive root express and extend, after 28d, add up adventive root incidence, record mean elements and observe root system quality (between rhizome callus a situation arises).Concrete data are in Table 1.
(6) test-tube seedling transplanting
Regeneration plant root agar is cleaned, transplanted in peat soil, vermiculite and perlite weight ratio and be: in the mixed-matrix of 1: 1: 1, cultivate, water spray moisturizing, added up transplanting survival rate after one month.Concrete data are in Table 1.
The condition of culture temperature of said process: 25 ℃ of daytimes, 20 ℃ of evenings, illumination 2000lx, illumination every day 16h.
embodiment 3
1, experiment material
Take masson pine then ripe cone seed germination germ free apical bud be explant, cone picks up from Ningming of Guangxi portiatree provenance construction masson pine seed production stand the good masson pine family of high yield fat Ningming No. 7.
2, experimental technique
(1) clump bud induction
Aseptic explant is inoculated into improvement MS+6-BA 4.0mgL -1+ NAA 0.1mgL -1+ sucrose 30000mgL -1+ agar 3500mgL -1medium on induce cultivation.Cultivate after 26d, average clump bud induction rate reaches 99.0%.
The basic composition of described modified MS medium is with embodiment 1.
(2) clump bud extends
The budlet that grows thickly inducing is transferred to improvement MS+AC 4000mgL -1+ sucrose 30000mgL -1+ agar 3500mgL -1in elongation medium, to promote the elongation growth of culture.Extend and cultivate 28d, the high 3-5cm of clump bud.
(3) subculture bud is cultivated
The simple bud of plant height 2-3cm in Multiple Buds is cut, be inoculated into modified MS medium+6-BA 2.5mgL -1+ KT0.5mgL -1+ NAA 0.25mgL -1+ sucrose 30000mgL -1+ agar 3500mgL -1on carry out subculture bud propagation 26d.
Shoot proliferation bud is put in step (2) elongation medium, to promote shoot proliferation bud to extend.Extend and cultivate 25d, the high 3-5cm of shoot proliferation bud.Elongation subculture bud is divided into after the long terminal bud of 2cm and the long band of 1cm lateral bud bud section, and repeating step (3) shoot proliferation and step (2) clump bud extend.Subculture is cultivated 5 times, average growth coefficient 6.6.
(4) root induction
The single subculture bud that cuts robust growth, semi-lignified, high 2-3cm, is inoculated into 1/2 modified MS medium+NAA 0.05mgL -1+ IBA 1.0mgL -1the former base of inducing adventitious root in+sucrose 2.0% medium.After induction 25d, root induction rate reaches 82.5%.
(5) adventitious root extension
To obviously there is the visible adventive root explant of naked eyes and proceed to 1/2 modified MS medium+AC 3500mgL -1in+sucrose 2.0% medium, promote adventive root express and extend, after 28d, add up adventive root incidence, record mean elements and observe root system quality (between rhizome callus a situation arises).Concrete data are in Table 1.
(6) test-tube seedling transplanting
Regeneration plant root agar is cleaned, transplanted in peat soil, vermiculite and perlite weight ratio and be: in the mixed-matrix of 1: 1: 1, cultivate, water spray moisturizing, added up transplanting survival rate after one month.Concrete data are in Table 1.
The condition of culture temperature of said process: 26 ℃ of daytimes, 19 ℃ of evenings, illumination 2000lx, illumination every day 16h.
In above embodiment 1 to 3, masson pine group training subculture blastogenesis root and transplant survival situation, as shown in Table 1 below.
Table 1 masson pine group training subculture blastogenesis root and transplant survival situation
From table 1 result, show, adopt the present invention that each family group training seedling subculture bud rooting rate of masson pine is reached more than 82.5%, the highest rooting rate reaches 90.6%, and more than the average root of hair 2-3 of every plant bar, and the callus of group training seedling base portion is less, between rhizome, connects closely.Transplant to after in peat soil, perlite, vermiculite mixed-matrix, regeneration plant well-grown, after one month, transplanting survival rate is more than 85.1%, and the highest rooting rate reaches 93.0%.The present invention has solved the problem of masson pine group training subculture blastogenesis root difficulty preferably, produces masson pine tissue culture regeneration plant group training subculture blastogenesis method for root is provided for batch production.

Claims (8)

1. one kind promotes masson pine group training subculture blastogenesis method for root, it is characterized in that: the two step rooting methods that comprise clump bud induction, the elongation of clump bud, subculture cultivation, root induction, adventitious root extension, test-tube seedling transplanting, obtain masson pine group training subculture blastogenesis offspring, its operating procedure is as follows:
(1) clump bud induction: by after masson pine seed sterilization, carry out seed germination under aseptic condition, then intercepting and being with the long hypocotylar germ free apical bud of 0.5 cm is explant, carries out clump bud induction in inducing culture I;
(2) clump bud extends: the Multiple Buds inducing is extended in elongation medium II to cultivation;
(3) subculture is cultivated: the single terminal bud that cuts elongation bud is inoculated in shoot proliferation medium III and is bred, obtain subculture bud I; Subculture bud I repeating step (2) is extended, singlely cut the subculture bud I of elongation and be divided into terminal bud and band lateral bud bud section, then in step (3) shoot proliferation medium, continue propagation and obtain subculture bud II, by subculture bud II repeating step (2) and step (3) again, obtain new proliferation and subculture bud; Repetitive cycling step (2) and (3), until obtain a large amount of enough culture of rootage subculture buds;
(4) root induction: the single subculture bud that cuts robust growth, semi-lignified, high 2-3 cm, be seeded in the 1/2 modified MS medium IV that contains methyl α-naphthyl acetate (NAA), indolebutyric acid (IBA), sucrose, adventitious root primordia 21-28 d is processed in induction;
(5) adventitious root extension: will there is the visible adventive root explant of naked eyes and proceed in the 1/2 modified MS medium V that contains active carbon (AC) or ascorbic acid (Vc), sucrose, and promote expression and the elongation of adventive root;
(6) test-tube seedling transplanting: the Transplantation of Regenerated Plantlets of having taken root is cultivated in peat soil, vermiculite and perlite (1: 1: 1) mixed-matrix to water spray moisturizing.
2. a kind of promotion masson pine group training subculture blastogenesis method for root according to claim 1, is characterized in that: described its material content of medium I is: 6-BA 2.5-4.0 mgL -1, NAA 0.05-0.1 mgL -1, sucrose 30000 mgL -1, acid hydrolyzed casein 500 mgL -1and modified MS medium.
3. a kind of promotion masson pine group training subculture blastogenesis method for root according to claim 1, is characterized in that: described its material content of medium II is: NAA 0.25-0.4 mgL -1or active carbon 3000-5000 mgL -1, sucrose 30000 mgL -1and modified MS medium.
4. a kind of promotion masson pine group training subculture blastogenesis method for root according to claim 1, is characterized in that: described its material content of medium III is: 6-BA 1.0-3.0 mgL -1, KT 0-1.0 mgL -1, NAA 0-0.3 mgL -1, sucrose 30000 mgL -1and modified MS medium.
5. a kind of promotion masson pine group training subculture blastogenesis method for root according to claim 1, is characterized in that: described its material content of medium IV is: NAA 0.05-0.1 mgL -1, IBA 1.0-2.5 mgL -1, sucrose 1.0-2.0% and 1/2 modified MS medium.
6. a kind of promotion masson pine group training subculture blastogenesis method for root according to claim 1, is characterized in that: described its material content of medium V is: AC 3000-5000 mgL -1or Vc 10-30 mgL -1, sucrose 2.0% and 1/2 modified MS medium.
7. a kind of promotion masson pine group training subculture blastogenesis method for root according to claim 1, is characterized in that: basic composition is of described modified MS medium: KNO 31650 mgL -1; NH 4nO 3600 mgL -1; CaCl 22H 2o 260 mgL -1; MgSO 47H 2o 370 mgL -1; Ca (NO 3) 24H 2o 400 mgL -1; KH 2pO 4170 mgL -1; MnSO 4h 2o 22.3 mgL -1; ZnSO 47H 2o 8.6 mgL -1; CuSO 45H 2o 0.025 mgL -1; H 3bO 36.2 mgL -1; Na 2moO 42H 2o 0.025 mgL -1; KI 0.83 mgL -1; CoCl 26H 2o 0.025 mgL -1; Cobastab 11.0 mgL -1; Cobastab 60.5 mgL -1; Nicotinic acid 0.5 mgL -1; Glycine 2.0 mgL -1; Inositol 200 mgL -1.
8. a kind of promotion masson pine group training subculture blastogenesis method for root according to claim 1, is characterized in that: described cultivation controlled condition is: cultivation temperature 22-26 ℃ on daytime, evening 16-20 ℃, illumination 1500-2000 lx, illumination every day 16 h.
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CN104756869A (en) * 2015-04-09 2015-07-08 广西壮族自治区林业科学研究院 Method for preparation of pinus massoniana lamb superior individual sterile explant and induction of initial bud
CN104756869B (en) * 2015-04-09 2016-04-20 广西壮族自治区林业科学研究院 The preparation of masson pine fine individual plant aseptic explant and initial bud inducement method
CN105359948A (en) * 2015-09-11 2016-03-02 中国林业科学研究院林业研究所 Hybrid larch micro-cuttage seedling growing method
CN105746288A (en) * 2016-04-15 2016-07-13 中国林业科学研究院热带林业研究所 Establishing method of pinus kesiya royle ex gordon cutting and breeding system
CN105746288B (en) * 2016-04-15 2019-05-28 中国林业科学研究院热带林业研究所 A kind of method for building up of cassie Asia pine cuttage nursery system
CN106688885A (en) * 2016-11-15 2017-05-24 邹少英 Method for quickly obtaining masson pine vegetative propagation seedlings for afforestation
CN108967195A (en) * 2018-08-07 2018-12-11 广西壮族自治区林业科学研究院 A kind of cultural method of rooting of masson pine tissue culture bud proliferation and rejuvenation
CN108967195B (en) * 2018-08-07 2021-07-02 广西壮族自治区林业科学研究院 Culture method for proliferation and rejuvenation of tissue culture subculture bud of pinus massoniana
CN111616050A (en) * 2020-05-19 2020-09-04 广西壮族自治区林业科学研究院 Method for improving rooting stability of superior tree subculture buds of high-yield masson pine
WO2021232507A1 (en) * 2020-05-19 2021-11-25 广西壮族自治区林业科学研究院 Method for improving rooting stability of subculture buds of high-fat-yield pinus massoniana superior tree
CN111616050B (en) * 2020-05-19 2022-11-15 广西壮族自治区林业科学研究院 Method for improving rooting stability of superior tree subculture buds of high-yield masson pine
CN111657150A (en) * 2020-07-24 2020-09-15 广西壮族自治区林业科学研究院 Method for improving rooting capacity of masson pine based on endogenous hormone

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