CN104130969A - Culture medium for increasing dry weight of cotton fibers and culture method thereof - Google Patents
Culture medium for increasing dry weight of cotton fibers and culture method thereof Download PDFInfo
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- CN104130969A CN104130969A CN201410365656.3A CN201410365656A CN104130969A CN 104130969 A CN104130969 A CN 104130969A CN 201410365656 A CN201410365656 A CN 201410365656A CN 104130969 A CN104130969 A CN 104130969A
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Abstract
The invention relates to the field of cotton fibers, and particularly relates to a culture medium for increasing the dry weight of cotton fibers and a culture method thereof. According to the culture medium for increasing the dry weight of the cotton fibers, a second component is added into a BT culture medium, wherein the second component is selected from any one of the following components: mannitol, methyl jasmonate, sucrose, and potassium chloride. The culture medium for increasing the dry weight of the cotton fibers adopts a cotton ovule in-vitro culture method, excludes the whole plant participation complexity, excludes changes of water, illumination, temperatures and the like under field conditions, and can artificially control the conditions for better studying on a development mechanism of the cotton fibers under the in-vitro condition; and the dry weight of the cotton fibers cultured through the culture medium is improved greatly, and a great significance is provided for improvement of the yield of cotton.
Description
Technical field
The present invention relates to cotton fiber field, in particular to a kind of substratum and cultural method thereof that increases cotton fiber dry weight.
Background technology
Cotton, as a kind of important cash crop and textile fiber material, is the important channel of increasing peasant income, is people's one of warming indispensable materials of living, and has important strategic position in national economy.Therefore cotton variety, how to cultivate high yield, high-quality always is the important goal of breeding men.
Cotton fibre is formed by ovule epidermic cell Fiber elongation, is substantially divided into differentiation projection, elongation, secondary wall thickening and the ripe four-stage that dewaters.Cotton fibre is the important component part of cotton boll, and fibrous quality and output are the coefficient results of genotype and ecological condition, be formed with the not same-action of different development stage to fibrous quality and output.The initial differentiation of cotton fibre occurs in 3-0DPA (daypostanthesis), is the beginning of cotton fiber development.The differentiation of this stage fiber cell is directly determining on cottonseed the radical of raw fiber, and its differentiation sooner or later directly affects the length of mature fibers, and finally affects lint yield; At elongate fiber, in the phase, the extension speed of fiber, elongating stage, rapid elongation phase etc. affect the length variations of fiber, and they are subject to the regulation and control of Endogenous Hormone Contents in Vitro and related enzyme activity in growth course.
Secondary wall thickening growth course is the critical period that fibre strength forms, and major effect fineness and the intensity of cotton fiber, can finally affect the performance of fiber strength; In Cotton fiber thickening development process, with fibrin deposition amount, increase, fiber dry-matter accumulation amount improves, also there is respective change in fibrous quality, under suitable ambient conditions, if secondary wall thickening is grown the time opening more early, and the time length is longer, the final fibre strength forming is just high, otherwise fibre strength is low.First the increase of fiber cell wall thickness is that fiber maturity improves constantly, and the increase of fiber dry weight constantly reduces micronaire value, and the continuous increase of fibre breakage specific tenacity is main relevant with fibrin deposition characteristic with Cultivar.
Cotton ovule culture system can be used for studying plant hormone and the impact of other material on Fibre Development as a good experimental system.Plant hormones regulators,gibberellins (GA3) and growth hormone (IAA) are the important factors that affects Study On Fiber Differentiation.In cotton ovule cultivation, GA3 can induce more fiber initiating cell, and GA3 and IAA can promote elongate fiber, and ABA suppresses elongate fiber.Secondary wall thickens phase IAA increases cellulose amount, and ABA plays opposite effect, and GA3 seems on the not impact of cellulosic amount.In rape element, cruel (BRs) can promote elongate fiber, initial all very important with elongation to fibrocellular differentiation, if process cotton germination with BRS biosynthesis inhibitor brassinazole2001 (Brz), causes there is no fibrocellular differentiation completely.Cotton steroids 5 alpha-reductase gene (GhDET2), this gene is the biosynthetic rate-limiting enzyme gene of BRs, further result of study shows that this gene is the highest at the expression level of cotton fiber cell rapid elongation phase, simultaneously this gene in Xuzhou 142 lint-free without the expression level in wadding mutant 0DPA ovule be only the same period wild-type ovule 1/5, illustrate that GhDET2 gene has vital role in the initial sum rapid elongation process of cotton fiber.
And Cotton in China yields poorly at present, can not meet domestic demand, improve Cotton in China fibrous quality, particularly adjusting cotton fiber dry weight becomes Cotton in China breeding key issue urgently to be resolved hurrily.
Summary of the invention
The object of the present invention is to provide a kind of substratum and cultural method thereof that increases cotton fiber dry weight, to solve the above problems.
A kind of substratum that increases cotton fiber dry weight is provided in an embodiment of the present invention, in BT substratum, has added second component, described second component is selected from any in following material: N.F,USP MANNITOL, methyl jasmonate, sucrose, Repone K.
Preferably, described second component is N.F,USP MANNITOL, and the content of described N.F,USP MANNITOL is 2-15g/L.
Preferably, the content of described N.F,USP MANNITOL is 5-8g/L.
Preferably, described second component is methyl jasmonate, and the content of described methyl jasmonate is 0.05-8 μ mol/L.
Preferably, the content of described methyl jasmonate is 0.5-5 μ mol/L.
Preferably, described second component is sucrose, and the content of described sucrose is 2-15g/L.
Preferably, the content of described sucrose is 5-10g/L.
Preferably, described second component is Repone K, and the content of described Repone K is 0.02-0.06mol/L.
Preferably, the content of described Repone K is 0.04-0.05mol/L.
Preferably, described BT substratum also contains indolylacetic acid and Plant hormones regulators,gibberellins; The addition of described indolylacetic acid is 8-12 μ mol/L, and the addition of described Plant hormones regulators,gibberellins is 4-6 μ mol/L.
The present invention also provides a kind of cultural method that increases cotton fiber dry weight, comprise the following steps: field cotton blooms and carries out selfing listing mark the day before yesterday, take away the ovule of the young bell of spending latter three days, then adopt the substratum of the increase cotton fiber dry weight described in right 1-9 any one to carry out in vitro dark cultivation; Culture temperature is 30 ℃ ± 2 ℃.
A kind of substratum that increases cotton fiber dry weight that the embodiment of the present invention provides, adopt cotton ovule isolated culture method, got rid of the complicacy that complete stool participates in, got rid of under field condition, the variation of moisture, illumination, temperature etc., control condition that can be artificial in vitro is better studied the development mechanism of cotton fibre; The dry weight of the cotton fiber obtaining by this culture medium culturing is greatly improved, and this raising to output of cotton is significant.
Embodiment
Below by specific embodiment, the present invention is described in further detail.
Embodiment 1
1.1 test sites are originally tested in January, 2014-May and are carried out in Hainan scientific research center of cotton biology National Key Laboratory of the Chinese Academy of Agriculture Science and Technologys Cotton Research Institute.
1.2 test materialss are brown 1-61, palm fibre 263, RT white cotton fiber, green cotton CC28, green cotton G88 for examination material, and all material Jun You the Chinese Academy of Agriculture Science and Technologys Cotton Research Institute's national middle term storehouse provides.All material has the annual selfing in this laboratory to preserve.The plant-growth regulator of experiment use has indolylacetic acid (IAA), Plant hormones regulators,gibberellins (GA3) and methyl jasmonate (MeJA) purchased from Aladdin.Repone K, sucrose and N.F,USP MANNITOL are domestic analytical pure.The chemical reagent that ovules culture in vitro is used is purchased from German Amresco company or U.S. Sigma company.
The configuration of BT substratum: BT culture medium prescription is as shown in table 1.By formula in material prepare after, pH is adjusted to 5.0, BT substratum used is divided in 100ml Erlenmeyer flask, every bottle adds 50ml substratum, through autoclaving (121 ℃, 20min), standby.
Table 1 BT culture medium prescription
1.3 test method
1.3.1 ovule is cultivated
When blooming the day before yesterday, field cotton carries out selfing listing mark; In 8:00--9:00 in the morning, take away and spend young age of latter three days and carry out ovules culture in vitro, the young bell of ovule first to peel off after bract and sepal again with 70% alcohol ovary sterilization 3～4min; Alcohol with 95% after sterilization infiltrates 2-3 second; Then after alcohol exposure lamp flame envelope, be put into the culture dish through sterilizing, after fray-out of flame, with tip tweezers, carefully separate ovule; Ovule is inoculated on the substratum of different treatment, is positioned in the culturing room of 30 ℃ ± 2 ℃ and secretly cultivates.Each processes minute 3 groups of cultivations, 3 bottles every group, 10～15 pieces of ovules of every bottle graft kind.
Better for ovule growth conditions, the parameter difference between the ovule of cultivation, apart from less, preferably, adds indolylacetic acid and Plant hormones regulators,gibberellins in described BT substratum; The addition of described indolylacetic acid is 8-12 μ mol/L, and the addition of described Plant hormones regulators,gibberellins is 4-6 μ mol/L.
1.3.2 the growth of ovule of observing BT culture medium culturing is dynamic
Observe DPC=5,10,15,20,25,30 (represent the number of days after cultivating, DPC=0 is that ovule is cultivated the same day) growth and dynamically, find, while cultivating 30 days, the fiber growth of ovule reaches stable maturity state.Therefore, select the cultivation ovule of 30 days to measure the dry weight of fiber.
When different culture media is cultivated, control medium is set, only adds the indolylacetic acid of corresponding content and the BT substratum of Plant hormones regulators,gibberellins.Wherein, before sucrose, Repone K and N.F,USP MANNITOL sterilizing, add; Methyl jasmonate adopts filtration sterilization, adds in the substratum after sterilizing after filtration sterilization.
Material in each test group comprises the several of brown 1-61, palm fibre 263, RT white cotton fiber, green cotton CC28, green cotton G88, and each processes minute 3 groups of cultivation, and 3 bottles every group, the kinds of culture medium of 10-15 piece of ovule employing of every bottle graft kind is as shown in test group 1-6.
Test group 1
Control group: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, the addition of Plant hormones regulators,gibberellins is 5 μ mol/L;
Group 1: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, N.F,USP MANNITOL in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of N.F,USP MANNITOL is 2g/L;
Group 2: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, N.F,USP MANNITOL in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of N.F,USP MANNITOL is 5g/L;
Group 3: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, N.F,USP MANNITOL in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of N.F,USP MANNITOL is 8g/L;
Group 4: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, N.F,USP MANNITOL in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of N.F,USP MANNITOL is 15g/L.
Test group 2
Control group: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, the addition of Plant hormones regulators,gibberellins is 5 μ mol/L;
Group 1: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, methyl jasmonate in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of methyl jasmonate is 0.05 μ mol/L;
Group 2: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, methyl jasmonate in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of methyl jasmonate is 0.5 μ mol/L;
Group 3: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, methyl jasmonate in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of methyl jasmonate is 5 μ mol/L;
Group 4: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, methyl jasmonate in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of methyl jasmonate is 8 μ mol/L.
Test group 3
Control group: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, the addition of Plant hormones regulators,gibberellins is 5 μ mol/L;
Group 1: ovules culture medium: add indolylacetic acid, Plant hormones regulators,gibberellins, sucrose in BT substratum, the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of sucrose is 2g/L;
Group 2: ovules culture medium: add indolylacetic acid, Plant hormones regulators,gibberellins, sucrose in BT substratum, the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of sucrose is 5g/L;
Group 3: ovules culture medium: add indolylacetic acid, Plant hormones regulators,gibberellins, sucrose in BT substratum, the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of sucrose is 10g/L;
Group 4: ovules culture medium: add indolylacetic acid, Plant hormones regulators,gibberellins, sucrose in BT substratum, the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of sucrose is 15g/L.
Test group 4
Control group: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, the addition of Plant hormones regulators,gibberellins is 5 μ mol/L;
Group 1: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, Repone K in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of Repone K is 0.02mol/L;
Group 2: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, Repone K in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of Repone K is 0.04mol/L;
Group 3: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, Repone K in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of Repone K is 0.05mol/L;
Group 4: ovules culture medium for to add indolylacetic acid, Plant hormones regulators,gibberellins, Repone K in BT substratum, and the addition of indolylacetic acid is 10 μ mol/L, and the addition of Plant hormones regulators,gibberellins is 5 μ mol/L, and the content of Repone K is 0.06mol/L.
Get at random ovule, with filter paper, surface liquid is blotted, peel off fiber, in 85 ℃ of left and right, fiber is dried to constant weight, weigh and obtain fiber dry weight.Wherein, the cotton of each kind is got at random respectively 20 ovules and measures fiber dry weight, averages.Data analysis to the fiber dry weight obtaining, the result obtaining is as shown in table 2-5.Wherein, rate of increase=(sample fiber dry weight-control group fiber dry weight)/control group fiber dry weight * 100%.
It should be noted that, a cotton has 30 ovules at least.
The fiber dry weight of each cotton variety in table 2 test group 1
As can be seen from Table 2, when the content of N.F,USP MANNITOL is 2-15g/L, its fiber dry weight all has increase, and wherein, when the content of N.F,USP MANNITOL is 2-5g/L, the rate of increase of fiber dry weight is higher.
The fiber dry weight of each cotton variety in table 3 test group 2
As can be seen from Table 3, when the content of methyl jasmonate is 0.05-8 μ mol/L, its fiber dry weight all has growth, and wherein, when the content of methyl jasmonate is 0.5-5 μ mol/L, its fiber dry weight rate of increase is higher.
The fiber dry weight of each cotton variety in table 4 test group 3
As can be seen from Table 4, when the content of sucrose is 2-15g/L, its fiber dry weight all has increase, and wherein, the content of sucrose is 5g/L, and the rate of increase of fiber dry weight is higher.
The fiber dry weight of each cotton variety in table 5 test group 4
As can be seen from Table 5, when the content of Repone K is 0.02-0.06mol/L, its fiber dry weight all has increase, and wherein, the content of Repone K is 0.04-0.05mol/L, and the rate of increase of fiber dry weight is higher.
The foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, for a person skilled in the art, the present invention can have various modifications and variations.Within the spirit and principles in the present invention all, any modification of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (10)
1. a substratum that increases cotton fiber dry weight, is characterized in that, in BT substratum, adds second component, and described second component is selected from any in following material: N.F,USP MANNITOL, methyl jasmonate, sucrose, Repone K.
2. the substratum of increase cotton fiber dry weight according to claim 1, is characterized in that, described second component is N.F,USP MANNITOL, and the content of described N.F,USP MANNITOL is 2-15g/L.
3. the substratum of increase cotton fiber dry weight according to claim 2, is characterized in that, the content of described N.F,USP MANNITOL is 5-8g/L.
4. the substratum of increase cotton fiber dry weight according to claim 1, is characterized in that, described second component is methyl jasmonate, and the content of described methyl jasmonate is 0.05-8 μ mol/L.
5. the substratum of increase cotton fiber dry weight according to claim 4, is characterized in that, the content of described methyl jasmonate is 0.5-5 μ mol/L.
6. the substratum of increase cotton fiber dry weight according to claim 1, is characterized in that, described second component is sucrose, and the content of described sucrose is 2-15g/L.
7. the substratum of increase cotton fiber dry weight according to claim 6, is characterized in that, the content of described sucrose is 5-10g/L.
8. the substratum of increase cotton fiber dry weight according to claim 1, is characterized in that, described second component is Repone K, and the content of described Repone K is 0.02-0.06mol/L.
9. according to the substratum of the increase cotton fiber dry weight described in claim 1-8 any one, it is characterized in that, described BT substratum also contains indolylacetic acid and Plant hormones regulators,gibberellins; The addition of described indolylacetic acid is 8-12 μ mol/L, and the addition of described Plant hormones regulators,gibberellins is 4-6 μ mol/L.
10. a cultural method that increases cotton fiber dry weight, it is characterized in that, comprise the following steps: field cotton blooms and carries out selfing listing mark the day before yesterday, take away the ovule of the young bell of spending latter three days, then adopt the substratum of the increase cotton fiber dry weight described in right 1-9 any one to carry out in vitro dark cultivation; Culture temperature is 30 ℃ ± 2 ℃.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20010018773A1 (en) * | 1995-02-21 | 2001-08-30 | Yoshihisa Kasukabe | Production of cotton fiber with improved fiber characteristics |
| CN103931494A (en) * | 2014-03-17 | 2014-07-23 | 安徽农业大学 | Colored cotton ovule in vitro culture method |
| CN104086314A (en) * | 2014-07-24 | 2014-10-08 | 中国农业科学院棉花研究所 | Culture medium for deepening color of brown cotton and culture method thereof |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20010018773A1 (en) * | 1995-02-21 | 2001-08-30 | Yoshihisa Kasukabe | Production of cotton fiber with improved fiber characteristics |
| CN103931494A (en) * | 2014-03-17 | 2014-07-23 | 安徽农业大学 | Colored cotton ovule in vitro culture method |
| CN104086314A (en) * | 2014-07-24 | 2014-10-08 | 中国农业科学院棉花研究所 | Culture medium for deepening color of brown cotton and culture method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 宋艳梅: "油菜素内酯对棉花纤维分化发育的影响及其调控基因bzr1的转化", 《中国优秀博硕士学位论文全文数据库（硕士）》 * |
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