CN1720802A - A biological sprout inhibitor and application thereof - Google Patents

Abstract

The invention relates to a biological anti-sprouting agent and its use, wherein the anti-sprouting agent is prepared from biological strains through the steps of enlarged culture and fermentation, solvent extraction, concentration, separation and purification. The bacterial strain has a systematic name of Botrytis cinerea, its active constituents include natural tyrosol and phenethylol wherein the contents are 25-30% and 22-25%.

Description

A kind of biological sprout inhibitor and application thereof

Technical field:

The present invention relates to a kind of biological sprout inhibitor and application thereof, belong to biological technical field.

Background technology:

Tobacco leaf is big in the western China cultivated area, and economic benefit is obvious.But after binding in the annual field management pruned, traditional craft removed the time-consuming fatigue of bud, and the labour drops into huge, and the wooden fork bud that long-time president again makes new advances behind the excision axillalry bud.In order to improve the tobacco leaf product yield, quality and minimizing labour drop into, China successively presses down bud Flumetralin (maleic Min) from buying abroad, Maleichdrazide (maleic hydrazide or bud enemy), remove bud logical (Pendimethalin) etc., these medicines are used for field management, bud inhibition effect has their own characteristics each, but price higher (more than 13 yuan/mu), and they all are chemical synthesis preparations, use this class chemosynthesis to press down the bud agent, in residual tobacco leaf of part chemical substance and the soil, to the influence and the residual toxicity of Flue-cured tobacco Quality, problems such as environment and soil pollution are not resolved as yet.

After axillalry bud takes place for vegetables and flowers, also be to adopt traditional craft to remove bud, not only time-consuming effect but also poor.

Summary of the invention:

The objective of the invention is to overcome the deficiencies in the prior art, and provide a kind of natural biological that tobacco, vegetables and flowers press down bud that is used for of nontoxic, free from environmental pollution, easy to use, efficient and cheap to press down the bud agent.

The present invention is achieved in that

One, natural biological presses down the preparation (preparation method is with the biofermentation method of existing conventional) of bud agent

Biological bacterial strain of the present invention was Hhp-1, and the classification called after Botrytis cinerea of bacterial strain Hhp-1 is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, deposit number: CGMCC NO 0357-1 on September 3rd, 1998.

A. from plant, gather Botrytis cinereapersr mycosis and poison sample, select the typical sample of fresh symptom, carry out surface sterilization, wash with aqua sterilisa, again with separate tissue or the direct spore method of shaking, carrying out monospore separates, purifying is cultivated, then according to indoor bacterial strain screening and technological process of production method, the choice biologically active is strong, (productive rate is more than 0.8 ‰ for the high microbial strain system of productive rate, wheat presses down the test of bud fast detecting, and extract is diluted to 1200 times of unspent bacterial strains), through relatively filtering out superior strain Hhp-1 repeatedly, make the metabolism productive rate reach 1.5 ‰-2.3 ‰, number in regular turn at last, formally bacterial strain Hhp-1 is inserted and boil to half-mature sterilization barley corn, put into-40 ℃～-60 ℃ refrigerators and preserve standby.

B. Hhp-1 biology bacterial strain is put into the PDA medium, 25 ℃ of cultivations 10-12 days, filtration then added solvent ethyl acetate extraction 3 times, adds desiccant dehydration, concentrating under reduced pressure after the extraction, gets natural extract crude product, and it is standby to be placed on the interior preservation of refrigerator.The content of active component crude benzene ethanol, p-hydroxyphenylethanol is respectively 25%--30% and 22%--25% in the natural extract crude product.

C. the extract crude product is adopted indoor inhibition wheat bud scale growth method, carry out giving birth to fast surveying screening: extract crude product to be measured is diluted 50 times in advance, 100 times, 200 times ... 1000 times ... series concentration 5ml moves into respectively in the beaker that is placed with the wheat seed of handling, be advisable just to be dipped into seed, the contrast clear water, each concentration repeats 3 times, put in 25 ℃ of incubators and cultivated 5-7 days, observing the young root young shoot has or not and is suppressed phenomenon, the record result, tentatively judge whether contain activity or other approximate active compositions in the extract crude product, it is strong to filter out activity, the bacterial strain Hhp-1 that productive rate is high, and the enlarged culture fermentation obtains the extraction crude product.Use TLC and column chromatography purification crude product again, adopt GC/MS, HPLC method to analyze, use modern IR, UV, HRMS, the 2DNMR technology is determined molecular formula and structural formula, last scientifically definite two kinds of natural active components that press down the bud agent, be respectively p-hydroxyphenylethanol, benzyl carbinol, their molecular weight, molecular formula, structural formula are:

A. the molecular weight of p-hydroxyphenylethanol is 139.17, and molecular formula is C ₈H ₁₀O ₂, structural formula is:

B. the molecular weight of benzyl carbinol is 122.17, and molecular formula is C ₈H ₁₀O, structural formula is:

Two, multiple spot field plot trial:

The present invention carried out the multiple spot field plot trial in continuous 3 years, test method and result following (is example with the tobacco leaf) in the vegetables and the flowers main producing region in important tobacco leaf producing regions such as Yuxi, Yunnan Province, Muller, Xun Dian, Luliang, river and mountain and suburb, Kunming:

1. the selection of vega: select growth neat, the height unanimity, kind identical (being all " the big gold dollar of safflower " cigarette kind), normally bind prune after, treat that axillalry bud has just revealed about 1 centimeter long of point, divides the experimental plot by the field layout drawing.

2. test medicine: the extract crude product that fermentation obtains, maleic Min, bud enemy, to remove bud logical.

3. test medicine concentration: according to experimental scheme, take by weighing each reagent agent respectively, dissolve with medical 95% earlier, add water and stir and be modulated into desired concn: the extract crude product is diluted to 50 times, 100 times, 200 times; Maleic Min (active ingredient 25.5%) is diluted to 300 times; Bud enemy (active ingredient 30.2%) is diluted to 50 times; Remove bud logical (active ingredient 33%) and be diluted to 100 times.The clear water contrast.Totally 7 processing (containing the clear water contrast), 4 repetitions amount to 28 sub-districts, and arrange with the field random packet 0.03 mu of every sub-district.

4. dispenser agent mode: select sunny day not have the phase on rain day, extract axillalry bud from top to bottom successively, drip every strain flue-cured tobacco consumption 1ml at the wound place's point soup 1-2 that extracts axillalry bud by the cigarette strain.

5. result of the test

Multiple spot field plot trial result

Test medicine	The extract crude product	Maleic Min, bud oppose, remove bud and lead to
			Press down bud rate %	93.5	89-92
Cost (unit/mu)	7-8	≥13
			Residual and side effect	Do not have	Exist

From above multiple spot field plot trial result as can be seen, though the domestic chemosynthesis maleic Min of promoting the use of, bud enemy, remove bud logical etc. press down the bud rate can be up to 92%, but they exist tangible problem to be jointly: influence the tobacco leaf metabolism, cause part tobacco leaf teratogenesis, blade is little, cause quality of tobacco and output to descend, when using the people there is excitant, sometimes also can cause the human allergy to react, in the residual tobacco leaf of part chemical substance, the smoker has been increased harm, in the residual soil, be unfavorable for follow-up plant growth, and the use cost price is higher.

This is natural also to can be used for other plant after pressing down the bud dilution agent, and as the bud that presses down of flowers, vegetables, the regulation and control crop growth improves quality, improves output.

The invention has the advantages that:

1, naturally press down the bud agent owing to be to gather, separate, cultivate, filter, extraction, concentrate and get from plant, whole production is processed the use safety non-toxic, is had no side effect, noresidue, free from environmental pollution;

2, production cost is low, and low price, cost per mu be 7-9 unit only;

3, can improve the output of tobacco, flowers and vegetables, improve quality, but bud is efficient on average can be reached more than 93%.

Embodiment:

1, this natural biological presses down the preparation of bud agent

The preparation method of (one) description is basic identical in the preparation method that natural biological presses down the bud agent and the summary of the invention part.Wherein:

The Botrytis cinerea persr mycosis of gathering is poisoned sample and is derived from the plants such as strawberry, acid Chinese quince, flowers, lettuce.

2, this natural application that presses down the bud agent

This is natural press down bud agent preparation after, can adopt conventional drop, cup to drench or the spray mode, reached the overgrowing of inhibition cigarette bud, received good effect.30-40 days field efficacy phase, should press down the bud agent and also can be used for other plant, as the bud that presses down of flowers, vegetables, the regulation and control crop growth improves quality, improves output.

Embodiment 1:(is used for tobacco and presses down bud)

Extract crude product (natural p-hydroxyphenylethanol and benzyl carbinol) 10.0 grams that fermentation obtains

(95%) 50 milliliter of medical ethanol

950 milliliters in water

This is the biotype tobacco germination inhibitor application liquid of 1000 milliliters of hydroxyl benzyl carbinols and benzyl carbinol 0.5%, after employing drop mode is used, presses down the bud rate and on average can reach 93%-94%.

Embodiment 2 (being used for potato)

Extract crude product (natural p-hydroxyphenylethanol and benzyl carbinol) 8.0 grams that fermentation obtains

(95%) 40 milliliter of medical ethanol

960 milliliters in water

This is the biotype vegetable sprouting inhibitor application liquid of 1000 milliliters of hydroxyl benzyl carbinols and benzyl carbinol 0.4%, and after employing spray mode was used, potato sprouting was suppressed more than 93%.

Embodiment 3 (being used for Kang Naixing, China rose)

Extract crude product (natural p-hydroxyphenylethanol and benzyl carbinol) 7.0 grams that fermentation obtains

(95%) 35 milliliter of medical ethanol

965 milliliters in water

This is that the flowers of 1000 milliliters of hydroxyl benzyl carbinols and benzyl carbinol 0.35% press down the bud agent and use liquid, adopt a cup pouring mode to use after, press down the bud rate and on average can reach 91%-92%.

Claims (2)

1, a kind of biological sprout inhibitor by microbial strains, through enlarged culture fermentation, solvent extraction, concentrate, obtain after the separation and purification operation, is characterized in that:

(1) microbial strains was Hhp-1, and the classification called after Botrytis cinerea of Hhp-1 bacterial strain is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, deposit number: CGMCC NO 0357-1 on September 3rd, 1998;

(2) active component of this biological sprout inhibitor is two kinds of natural p-hydroxyphenylethanol and benzyl carbinols, wherein:

A. the content of crude benzene ethanol, p-hydroxyphenylethanol is respectively 25%--30% and 22%--25% in the natural extract crude product;

B. the molecular weight of p-hydroxyphenylethanol is 139.17, and molecular formula is C ₈H ₁₀O ₂, structural formula is:

C. the molecular weight of benzyl carbinol is 122.17, and molecular formula is C ₈H ₁₀O, structural formula is:

2, the said biological sprout inhibitor of claim 1 can be used as tobacco, vegetables and flowers and presses down bud agent application.