201010598 .…尤％:舞纖_^. 【發明所屬之技術領域】 本發明係關於微生物肥料之種子包埋方法及其產物。 【先前技術】 習常以環境中之資源用於農作物生產上，如將有益作物生長之微生物 接種到種子或施用於幼苗與土壤上，可增加植物營養要素之供應、提高土 Ο 壞中養分之有效性、增進根系之生長與養分之吸收、保護根系及增進抗逆 境能力等用途。故’利用微生物施予作物之產品又稱為「微生物肥料」。微 生物肥料之功能： (一）固氮作用：固氮菌包括共生、附著及游離三類菌，係將空氣中 的氮素固定為氨，轉變成作物可以利⑽氮源，固氮作用可減少氮肥的施 用，如此可減少為製造化學氮肥所消耗的石油能源，也可減緩土壞由於施 用化學肥料造成酸化的效應。 ® (—）；谷解作用.土壤中存有許多作物不能利用的不溶性營養元素， 如碟_、魏鐵等，需靠溶磷菌將磷溶解後才能被利用，因此，這類菌 具有提供作物營養的功能，減少化學肥料的_，例如溶韻。 (三） 增進根系、營養吸收及生長的作肖：微生物肥料中有増進根系營 養吸收與生長的_ ’增加根系吸收能力及表面積，即可減少化學肥料的 施用’提高土壤中的營養供應效率，如菌根菌。 (四） 促進有機物之分解：A分子之有機物需經由微生物分泌酵素分 201010598 0 解成較小的分子或無機物，才能被植物利用。 (五）增加植物對逆境的抗性與病原菌之拮抗：微生物的部分分泌物 質對其他生物之生存具有抑制作用，此種微生物可使土壤或植物體中病原 菌減少甚或死亡，減少病害發生，降低農藥之使用。 植物病害之生物防治即是利用自然界的拮抗微生物活性及所能展現的 拮抗能力，有效降低病源菌數目或活性，以減緩病害發生速率，達成病害 防治功效。早期學者研究有益於作物生育的微生物，大多侷限於固氮菌溶 φ磷菌的篩選，並以探討微生物與作物間的交互作用及相關代謝為主。農作 物及果樹等經濟作物中發現的内生菌已達745種（隸屬於9屬），這些内生 細菌大多為土壞微生物種類腸桿菌屬（办、假單胞菌屬 (尸此―/^^)、枯草桿菌屬（价等三屬，是最具有潛力的生物防 治菌種，不僅可以有效防治作物病害，並可促進植物生長。 蕈狀芽孢桿菌（5沉zY/z/jmyodes)屬革蘭氏陽性，好氣或兼性厭氧性， 具周生鞭毛及產生内生孢子，普遍存在於土壤、植物體體表與組織内。型 Ο 態為短桿，長5微米，寬800奈米。由於該菌孢子可在自然界逆境下存活， 同時在產孢過程中，會分泌具有抑菌作用之抗生物質，具有防治植物病害 之功效（如：番茄萎凋病），另，文獻中記載枯草桿菌屬助）細菌除 能產生抗菌物質外’亦具促進作物生長與提高產量的功效。 然而，微生物肥料係活的微生物，非肉眼所能見，其菌數、活性與肥 效之良否實非一般農民所能辨識，且培養微生物施用之方法、技術門檻高， 致使該方法及應用不易普及；且如何將正確之菌種於適當時間、適當環境 201010598 * 施予適當之作物，以促使微生物及作物間達到協同共生之益，亟需經驗及 高技術門權。另，微生物肥料若能替代部分化學肥料之施用，舒減化學肥 料不當使用對環境之衝擊，對農業永續發展當有很大的助益。 由此可見’上述習用方法及物品仍有諸多缺失，實非一良善之設計者， 而亟待加以改良。 種子披衣技術最初應用於穀類種子上，目前種子彼衣技術已被廣泛應 用而成為「種子工業」’因其加工處理方法不同大致區分為三種類型包括： ❿一般粉衣（seed coat) ’膜衣處理（film coating)及改變種子的形狀、大小、 重量以配合精密的機械播種的造粒處理（seedpelleting)等。該三種處理方 法於實際應用生產時可獨立分開作業，可因作物類型、栽種模式、環境需 求結合不同披衣處理，讓種子不但易於機械操作，更具備制病、防蟲、抗 逆境、促進生長等附加價值》 種子披衣技術能受到許多先進國家重視，投入大量的研發人力與經 費’主要原因係其利於機械播種外，更可在批衣過程中加入藥劑、肥料、 _植物生長觸織有益微生物等，可有效㈣苗期病蟲害及促進生長；另 搭配不同物理、化學特性的披衣物質，使作物在不㈣土壤條件下能促進 種子之發芽或調控其發芽的時間。 是以，本案發明人鑑於上述習用微生物肥料所衍生的各項缺點，乃亟 思加以改良績，並經多年苦心孤錄心研究後，終於成功研發完成本件 促進植物生長及抵抗病源菌感染之種子包埋方法及其產物。 7 201010598 ， 【發明内容】 本發明之目的即在於提供一種促進植物生長及抵抗病源菌感染之種子 包埋方法’該方法藉由微生物包埋植物種子，以促進植物生長及抵抗病原 菌感染。 本發明之次一目的係在於提供一種促進植物生長及抵抗病源菌感染之 種子包埋方法所得產物，該產物係藉由微生物包覆植物種子，促使該植物 可抵抗病原菌感染及促進該植物之生長。 可達成上述發明目的之一種促進植物生長及抵抗病源菌感染之種子包 埋方法及其產物，包括有： 蕈狀芽抱桿菌（Bacinus! mycodies) ·, 蕈狀芽孢桿菌（5acz7/⑽培養，包括將菌落接種於平板上，. 培養約2天後，刮取單一菌落接種於燒瓶内，振盪培養數天；然後，將燒 瓶培養物接種於酸酵槽培養基内，振蘯培養數天，即得蕈狀芽孢桿菌發酵 液，再以連績式離心機進行分離，即可得菌泥及懸浮液； Φ 膜衣劑； 臈衣劑製備，包括將上述所得之菌泥加入膜衣劑中混勻得一混合物 料，並將該混合物料復水配製成懸浮液備用之； 載體，包括蔬菜種子，如番小白菜、青江菜、油菜、空心菜杏菜戋 蘇子等。 低溫低濕流動層包埋方法，包括將上述載體與上述膜衣劑懸浮液，及 適當添加劑置入流動層乾燥機，以低溫、低濕條件下進行種子包埋，即可 201010598 取出，以得到經蕈狀芽孢桿菌包埋之種子。 【實施方式】 冬發明之實施例所用之蕈狀芽孢桿菌（Bacmusmyc〇dies，BM)係得自 寄存於中華民國台灣省新竹市食品工業研究所_保存中心的蕈狀芽孢桿 菌 BCRC10044。 蕈狀芽孢桿菌（如勤寧0fe)培養，包括將菌落接種於平板上， 於適當溫度如15·37Υ，（較佳者周溫約3〇。〇下培養約2天後，麻單一菌 落接種於燒瓶内’訂列培養基，在約3〇Τ，振贱率1(M2Gipm (較佳者 為120 _之下減培養數天；然後，將燒瓶培養物接種於酸酵槽培養基(同 燒瓶培養基)内，在15-寶，（雛者周溫約取），漏G 81 G公斤坪方 么为，以75 rpm攪拌速度，以0.01-1.5 WM通氣速率通入空氣，或空氣與 氧氣，二氧化碳或氮氣的混合物，較佳者空氣，培養約3天，即得蕈狀芽 孢桿菌發酵液，再以連續式離心機(轉速為16〇〇〇 φιη)進行分離，即可得菌 泥及懸浮液； 該蕈狀芽孢桿菌之培養基配方如下; 培養基纪方 含量(重晉 0.01 〜5 0·01 〜2 0.0001〜0.2 0.01~ 10 0.001 〜2 、硫酸鐵等； 綜合性碳氮源 動植物來源蛋白及其水解物 無機鹽類 糖類 酵母或麥芽抽出物(粉、膏） 其中該無機鹽類可為硫酸鎮、磷酸氫二鉀 201010598 其中該糖類可為葡萄糖、蔗糖、 、果糖、麥芽糖等；201010598 .... 尤%:舞纤_^. [Technical Field of the Invention] The present invention relates to a seed embedding method of microbial fertilizer and a product thereof. [Prior Art] The use of resources in the environment for crop production, such as inoculation of beneficial crops to seeds or application to seedlings and soil, can increase the supply of plant nutrient elements and improve the soil nutrients. Effectiveness, increase root growth and nutrient absorption, protect roots and enhance resistance to stress. Therefore, products that use microbes to give crops are also called "microbial fertilizers." The function of microbial fertilizer: (1) Nitrogen fixation: Nitrogen-fixing bacteria include symbiotic, attached and free three types of bacteria, which fix nitrogen in the air to ammonia, and convert to crops to benefit (10) nitrogen source. Nitrogen fixation can reduce the application of nitrogen fertilizer. This can reduce the petroleum energy consumed for the manufacture of chemical nitrogen fertilizers, and can also alleviate the effect of acidification caused by the application of chemical fertilizers. ® (-); trough solution. There are many insoluble nutrients in the soil that cannot be used by crops, such as dish _, Wei iron, etc., which need to be dissolved by phosphorus solubilizing bacteria before they can be used. Therefore, these bacteria have been provided. The function of crop nutrition reduces the amount of chemical fertilizers, such as the rhyme. (3) Enhancing the root system, nutrient absorption and growth: microbial fertilizers have the roots of nutrient absorption and growth _ 'increasing root absorption capacity and surface area, which can reduce the application of chemical fertilizers' to improve the efficiency of nutrient supply in the soil, Such as mycorrhizal fungi. (4) Promoting the decomposition of organic matter: The organic matter of molecule A needs to be decomposed into micro-organisms or inorganic substances through microbial secretory enzymes 201010598 0 to be used by plants. (5) Increasing plant resistance to stress and antagonism of pathogens: Some secreted substances of microorganisms have an inhibitory effect on the survival of other organisms. Such microorganisms can reduce or even kill pathogenic bacteria in soil or plants, reduce disease occurrence, and reduce pesticides. Use. The biological control of plant diseases is to use the antagonistic microbial activity of nature and the antagonistic ability that can be exhibited, and effectively reduce the number or activity of pathogenic bacteria, so as to slow down the rate of disease occurrence and achieve disease control effects. Early scholars studied microorganisms that are beneficial to crop growth, and most of them were limited to the screening of nitrogen-fixing bacteria, and to explore the interaction between microorganisms and crops and related metabolism. There are 745 endophytic bacteria (subordinate to 9 genera) found in cash crops such as crops and fruit trees. Most of these endophytic bacteria are indigenous microorganisms of the genus Enterobacter (Pseudomonas, Pseudomonas). ^), Bacillus subtilis (prices and other three genera, is the most promising biological control species, not only can effectively control crop diseases, and can promote plant growth. Bacillus licheniformis (5 sink zY / z / jmyodes) leather Lang's positive, aerobic or facultative anaerobic, with peripheral flagellum and endophytic spores, commonly found in soil, plant body surface and tissues. The type is short, 5 microns long and 800 nanometers wide. Because the spores can survive in the natural environment, and in the sporulation process, they will secrete antibacterial substances with antibacterial action, which have the effect of controlling plant diseases (such as tomato wilt). In addition, the literature describes Bacillus subtilis. In addition to the ability to produce antibacterial substances, bacteria also have the effect of promoting crop growth and increasing yield. However, microbial fertilizers are living microorganisms that are not visible to the naked eye, and the number of bacteria, activity and fertilizer efficiency are not ordinary. It can be identified by the people, and the methods and techniques for cultivating microbial application are high, which makes the method and application difficult to popularize; and how to plant the correct strain at the appropriate time and appropriate environment 201010598 * Apply appropriate crops to promote microorganisms and crops To achieve the synergy of symbiosis, there is no need for experience and high-tech rights. In addition, if microbial fertilizer can replace the application of some chemical fertilizers, it will greatly reduce the impact of improper use of chemical fertilizers on the environment, and it will have a great impact on the sustainable development of agriculture. It can be seen that 'the above-mentioned methods and articles still have many defects, which are not a good designer, but need to be improved. Seed dressing technology was first applied to cereal seeds, and now the seed coating technology has been widely used. The "seed industry" is roughly divided into three types due to its processing methods: se general coat "film coating" and changing the shape, size and weight of seeds to match the precision Mechanical seeding granulation treatment (seedpelleting), etc. The three treatment methods can be independently divided in practical application production. The work can be combined with different shawls for crop type, planting mode and environmental needs, so that the seeds are not only easy to mechanically operate, but also have added value such as disease, insect, stress, and growth. Seed dressing technology can be advanced. The state attaches great importance to investing a large amount of R&D manpower and funding. The main reason is that it is beneficial to mechanical sowing. It can also add chemicals, fertilizers, _ plant growth, woven and beneficial microorganisms in the process of coating, which can effectively (4) seedling pests and diseases and promote growth. Another combination of different physical and chemical properties of the cloak material allows the crop to promote the germination of the seed or regulate the germination time under the soil conditions. Therefore, the inventors of the present invention have various shortcomings derived from the above-mentioned conventional microbial fertilizer. Nai Sisi improved his performance, and after years of painstaking research, he finally succeeded in researching and developing this seed embedding method and its products to promote plant growth and resistance to pathogenic bacteria infection. 7 201010598, SUMMARY OF THE INVENTION An object of the present invention is to provide a seed embedding method for promoting plant growth and resistance to pathogenic bacteria infection. The method embeds plant seeds by microorganisms to promote plant growth and resistance to pathogenic bacteria infection. A second object of the present invention is to provide a product obtained by a seed embedding method for promoting plant growth and resistance to pathogenic bacteria, which is to coat a plant seed with a microorganism to promote the plant to resist infection of the pathogen and promote the growth of the plant. . A seed embedding method and a product thereof for promoting plant growth and resistance to pathogenic bacteria infection, including: Bacinus! mycodies, Bacillus licheniformis (5acz7/(10) culture, including The colonies were inoculated on the plate, and after about 2 days of culture, a single colony was scraped and inoculated into the flask, and cultured for several days with shaking; then, the flask culture was inoculated into the acid fermentation tank medium, and shaken for several days to obtain The fermentation broth of Bacillus licheniformis is separated by a continuous centrifuge to obtain the slime and suspension; Φ film coating agent; preparation of the coating agent, including adding the above-mentioned bacterial sludge to the film coating agent and mixing A mixture of materials is obtained, and the mixture is reconstituted into a suspension for use; the carrier comprises vegetable seeds, such as Chinese cabbage, Qingjiang, rape, and spinach apricot, etc. Low temperature and low humidity flow layer embedding. The method comprises the steps of: placing the above carrier and the above-mentioned film coating agent suspension, and appropriate additives in a fluidized bed dryer, and performing seed embedding under low temperature and low humidity conditions, and then taking out the 201010598, so as to obtain Seeds embedded in Bacillus licheniformis. [Embodiment] The Bacmus myc〇dies (BM) used in the examples of the winter invention was obtained from the Food Industry Research Institute of Hsinchu City, Taiwan Province, Republic of China_Save Center Bacillus licheniformis BCRC10044. Culture of Bacillus licheniformis (such as Qinning 0fe), including inoculating the colony on a plate at a suitable temperature such as 15.37 Υ, (preferably, the peripheral temperature is about 3 〇. After the day, a single colony of inoculation was inoculated into the flask to set the medium at about 3 Torr, and the vibrating rate was 1 (M2Gipm (preferably 120 _ minus culture for several days; then, the flask culture was inoculated with acid) In the fermentation tank medium (with the flask medium), in the 15-bao, (the chick's weekly temperature is about to take), the leakage G 81 G kg is the square, at 75 rpm, and the air is introduced at a ventilation rate of 0.01-1.5 WM. , or a mixture of air and oxygen, carbon dioxide or nitrogen, preferably air, cultured for about 3 days, the fermentation broth of Bacillus licheniformis, and then separated by a continuous centrifuge (rotation speed of 16 〇〇〇 φιη), ie Bacterial slime and suspension; The culture medium of Bacillus is as follows; the content of the culture medium (re-recognition 0.01 ~ 5 0 · 01 ~ 2 0.0001 ~ 0.2 0.01 ~ 10 0.001 ~ 2, iron sulfate, etc.; comprehensive carbon and nitrogen source of animal and plant origin protein and its hydrolyzate inorganic salt The sugar-like yeast or malt extract (powder, paste) wherein the inorganic salt may be sulfuric acid town, dipotassium hydrogen phosphate 201010598, wherein the sugar may be glucose, sucrose, fructose, maltose, etc.;
其中該動馳來碰自及其轉物輕自物ept〇ne)。 黃豆 膜衣劑製備’包括將上述所得之献與_劑以適當比娜勻並復水 配製成膜衣劑懸浮液備用之，此即為 包含但不限於玉米殿粉、脫脂奶粉、 此即為覆蓋載體之膜衣；其中該赋形劑成分 奶粉、乳糖、糊精、聚糊精、環狀糊精、甲 雜基纖維素、殿粉或硬脂酸納，較佳者為玉米殿粉；其中菌泥之重量百分比 佔膜衣麵浮絲重4之25 wt% ;其㈣形狀重量百姐侧衣劑懸浮 液總重量之0.5-5 Wt%，較佳者為2 wt0/〇。 載體’包括蔬菜種子，如番小白菜、青江菜、油菜、空心菜、杏菜或 茄子等。 低溫低濕流動層包埋方法，包括將上述載體與上述膜衣劑懸浮液，置 入流動層乾燥機’以入風溫度控制24-28°C，槽内控制溼度約15%以下，膜 • 衣劑懸浮液的進料速度為1〇 L/hr，待膜衣劑懸浮液進料完畢後，含水率為 5%以下時，即可取出，以得到經蕈狀芽孢桿菌包埋之種子，故，藉由上述 包埋方法得將膜衣劑均勻包覆於種子表面，其中該膜衣劑係由25wt%蕈狀 芽孢桿菌及0.5-5 wt%賦形劑組成》 本發明係以下面的實施例予以示範闡明，但本發明不受下述實施例所 限制。 資施例一蕈狀芽孢桿菌之培養 201010598 * 本發明所用蕈狀芽抱桿菌⑽，bm)係得自寄存於中華 民國台灣省新竹市食品工業研究所菌種保存中心，其寄存編號為 BCRC10044。培養方法如下： 菌絲艚餡妷： 為寄存於食品工業研究所的菌株BCRC10044。 平板培卷： 挑選單一菌落以以劃線法接種於平板上接種於平板上，使用胰化大豆瓊 _ 脂培養基(Tiyptic soy agar，TSA培養基，購於Merck)，於30。0：下培養約2 天。 燒瓶培奉： 刮取平板上的單一菌落接種於燒瓶内，用下列培養基，在約3〇〇c下， 於搖動機上以振盪速率120 rpm振盪培養3天； 培養基配方： 缝 蔗糖 蛋白腺(peptone> 含量ί重量 6 0.5 0.5 1 酵母抽出物、粉、膏 玉米搬粉 黃豆粉 碟酸氫二# 1 硫酸鎂 0.1 0.1Among them, the move is coming from its own light and the object is ept〇ne). The preparation of the soybean film coating agent comprises the following steps: preparing the film coating agent suspension with appropriate ratio and rehydration, which is included, but not limited to, corn house powder, skimmed milk powder, which is a film coating covering the carrier; wherein the excipient component is milk powder, lactose, dextrin, polydextrin, cyclodextrin, methane cellulose, temple powder or sodium stearate, preferably corn powder; The weight percentage of the slime is 45% by weight of the floating surface of the film, and the weight of the (4) shape weight is 0.5-5 Wt% of the total weight of the side dressing agent suspension, preferably 2 wt0/〇. The carrier 'includes vegetable seeds such as Chinese cabbage, Qingjiang, rape, spinach, apricot or eggplant. The low-temperature low-humidity flow layer embedding method comprises the steps of: placing the above-mentioned carrier and the above-mentioned film coating agent suspension into a fluidized bed dryer to control the inlet air temperature by 24-28 ° C, and controlling the humidity in the tank to be less than 15%, and the membrane The feed rate of the coating suspension is 1 〇L/hr, and after the film coating agent suspension is completed, when the water content is 5% or less, it can be taken out to obtain the seed embedded by Bacillus licheniformis. Therefore, the film coating agent is uniformly coated on the surface of the seed by the above embedding method, wherein the film coating agent is composed of 25 wt% of Bacillus licheniformis and 0.5-5 wt% of the excipient. The examples are exemplified, but the invention is not limited by the following examples. The cultivation method of Bacillus licheniformis 201010598 * The Bacillus licheniformis (10), bm) used in the present invention was obtained from the Stem Storage Center of the Food Industry Research Institute of Hsinchu City, Taiwan Province of the Republic of China, and its registration number is BCRC10044. The culture method is as follows: Mycelium 艚 stuffing: It is a strain BCRC10044 deposited in the Food Industry Research Institute. Plate culture: A single colony was selected and inoculated on a plate by streaking on a plate, and cultured on a plate using a tryptic soy agar medium (Tiytic soy agar, TSA medium, purchased from Merck) at about 30. 2 days. Flask culture: A single colony on the plate was scraped and inoculated into the flask, and cultured for 3 days at a shaking rate of 120 rpm on a shaker at about 3 ° C with the following medium; Medium formula: Sewn sucrose protein gland ( Peptone> Content ί Weight 6 0.5 0.5 1 Yeast extract, powder, cream corn powder, soy bean powder, acid hydrogen II # 1 magnesium sulfate 0.1 0.1
培養基同上，將燒瓶培養液接種於發酵槽培養基内，槽壓控制 〇.8-l.〇kg/cm2 ’通氣量〇J5 VVM，授拌轉速75㈣，培養3天後即可獲得 201010598 • 蕈狀芽孢桿菌發酵液，發酵液以連續式離心機（轉速為l6000lpm)進行分 離，即得菌泥及懸浮液。 結果： 2000公升之發酵槽培養基，經發酵後可獲得菌泥35公斤（濕重），菌 數為 l.OxlO11 cfii/克。 實施例二低溫低濕流動層包埋方法 膜衣劑絮備： 取4公斤實施例-所得之菌泥與320克之玉米殿粉混勻，並復水至16 公斤配製颜衣麵浮液，即微覆謂衣。其巾闕泥之重量百分 比佔膜衣劑懸浮液總重量之25 wt% ;其中該玉米婦之重量百分比倾衣 劑懸浮液總重量之2 wt%。 載體： 以l〇kg蔬菜種子（番小白菜、青江菜、油菜、空心菜、杏菜或莊子等） 為載體。 惠遥低滿流動層包埋方法： 將上述載艘與上述膜衣麵浮液置人市售之流動層乾賴，藉由反覆 噴塗該膜衣讎浮祕紐表面，經低溫風乾，使膜衣網勻包覆載體， 簡述其下：入風溫度控制24_28〇c，槽内控制渔度約⑽以下，膜衣劑懸浮 液的進料速度為10 L/hr，賊鋪祕液餅完畢後，待含轉為州以下 時即可取出’以得到含實施例一成分之包埋種子，下稱bm種子包埋組。 12 201010598 包埋種子之含菌數測詁： 測試方法為所屬技藝領域具有通常知識者習知之菌量測試方法，簡述 如下：取絲包㈣子料，將㈣種子置於液態培養射減以取得菌 液’再將g驗序列轉後，塗於培養基進行培養，以計算每粒包埋種子 之平均含菌量。 結果： 包埋後之種子其蕈狀芽孢桿菌菌數可達lxl09cfij/粒。 ® 實施例三溫室應用情形 試驗方法： 將對照組（未包埋）及腿種子包埋組（種子分別為小白菜、青江菜、 油菜、空心菜及杏菜之種子，經實施例二之包埋方法所得之包埋種子），每 組種植40株進行試驗。試驗進行三星期後，測量蔬菜之株高、葉片數、根 長，及植株乾重，植株乾重部分以8(^C烘箱烘乾植物4小時後測量之。 結果： 如表一所示’ BM種子包埋組的各組生長參數比對照組(未包埋組)佳， 該生長參數如株高、葉片數及植物乾重，尤其在植物乾重部分的表現高出 35%。故’種子包埋對於小白菜的生長勢有正面影響。 表一小白菜種子包埋之生長狀態統計___ __對照組_BM種子包埋紐 株高（公分） 24.4±2.12 27±1.54 葉片數（片） 7.1+0.56 7.6±0.69 13 201010598 ' ’ 根長（公分） 32.4±2.33 植株乾重（克） 26±2.06 26.4±2·36 35 士 1.92 2.青江菜 如表二所示，ΒΜ種子包埋組中之根長及植株乾 種子包埋對於青江菜的生長勢有正面影響。 重皆較對照組佳，故， 表二青江菜種子包埋之生長狀態統計The medium is the same as above, and the flask culture solution is inoculated into the fermentation tank medium, and the tank pressure is controlled to be 88-l.〇kg/cm2 'ventilation volume 〇J5 VVM, and the mixing speed is 75 (four). After 3 days of culture, the product can be obtained 201010598. The fermentation broth of Bacillus, the fermentation broth is separated by a continuous centrifuge (rotation speed of l6000 lpm), that is, the bacterial sludge and the suspension are obtained. RESULTS: The fermentation broth of 2000 liters was fermented to obtain 35 kg (wet weight) of bacteria sludge, and the number of bacteria was l.OxlO11 cfii/g. Example 2 Low-temperature and low-humidity flow layer embedding method Film coating agent preparation: Take 4 kg of the sample-derived slime and 320 g of corn house powder, and rehydrate to 16 kg to prepare the face-covering float, ie Micro cover is called clothing. The weight percent of the towel mud is 25 wt% of the total weight of the film coating suspension; wherein the weight percentage of the corn kernel is 2 wt% of the total weight of the coating agent suspension. Carrier: l〇kg vegetable seeds (Pan Xiaocai, Qingjiang, rape, spinach, apricot or Zhuangzi) are used as carriers. Huiyao low-full-flow layer embedding method: The above-mentioned carrier and the above-mentioned film-coating floating liquid are placed on the commercially available flowing layer, and the film is sprayed by re-spraying the surface of the film coating The clothing net is evenly coated with the carrier, and the following is briefly described: the inlet air temperature is controlled 24_28〇c, the controlled fishing degree is below (10), the film coating agent suspension is fed at 10 L/hr, and the thief shop secret cake is finished. After that, when the content is changed to the state below, it can be taken out to obtain the embedded seed containing the component of the first embodiment, which is hereinafter referred to as the bm seed embedding group. 12 201010598 The number of bacteria contained in the embedded seeds: The test method is a method for testing the amount of bacteria known to those skilled in the art, and is briefly described as follows: taking the silk package (4), and placing the seeds in the liquid culture The bacterial liquid was obtained, and then the g-sequence was transferred, and the medium was cultured to calculate the average bacterial amount of each embedded seed. Results: The number of Bacillus licheniformis seeds after embedding can reach lxl09cfij/grain. ® Example 3 Greenhouse Application Situation Test Method: The control group (unembedded) and leg seed embedding group (seeds are seeds of Chinese cabbage, Qingjiang, rape, water spinach and apricot, respectively, and the embedding method of the second embodiment) The resulting embedded seeds were tested in groups of 40 plants. Three weeks after the test, the plant height, the number of leaves, the root length, and the dry weight of the plants were measured, and the dry weight of the plants was measured after drying the plants for 4 hours in an oven (Results: as shown in Table 1) The growth parameters of each group in the BM seed-embedded group were better than those in the control group (un-embedded group). The growth parameters such as plant height, number of leaves and dry weight of the plant, especially in the dry weight of the plant, were 35% higher. Seed embedding has a positive effect on the growth potential of pakchoi. Table 1 Statistics on the growth status of pakchoi seed embedding ___ __ control group _BM seed embedding plant height (cm) 24.4 ± 2.12 27 ± 1.54 number of leaves ( 7.1+0.56 7.6±0.69 13 201010598 ' ' Root length (cm) 32.4±2.33 Plant dry weight (g) 26±2.06 26.4±2·36 35 ± 1.92 2. Qingjiang cuisine as shown in Table 2, ΒΜ seed package The root length in the buried group and the dry seed embedding of the plant had a positive effect on the growth potential of Qingjiang cuisine. The weight was better than that of the control group. Therefore, the growth status of the seed embedding of Qingjiangcai was shown in Table 2.
包埋組 24.3+1.47 7·6 士 0.58 32.8±2.14 24±1.〇4 對照組 株高（公分） 2611.55 ®葉片數（片） 7.5±0.87 根長（公分） 25.2±1.44 植株乾重（克） 20+1.69 3.油菜 數高較對照參 是土」φ菜種子包埋之生長狀熊欲計 -- -對照組 φ 株高（公分） 35_8±2.13 葉片數（片） 5.6±0.25 根長（公分） 30.4±1.35 (克） 3011.01 包埋組 37.3+1.54 5.3+0.36 25.5±1.24 28±1.60 4.空心菜 如表四所示，BM種子包埋組中除了根長的差異性不大之外，其他生長 參數都顯示以BM將種子包埋後對於空心菜生長勢較佳。 表四空心菜種子包埋之生長狀態統計 201010598 對照組 BM種子包埋組 株高（公分） 66.7±2.17 71.5±2.22 葉片數（片） 32±2.03 34.7±2.57 根長（公分） 30.9±1.39 28.2±1.〇1 植株乾重（克） 38±1.32 4511.63 5·杏菜 如表五所不，杏菜之ΒΜ種子包埋組中除根長外，其餘生長參數皆佳 於對照組。故證實以ΒΜ將種子包埋對於杏菜之生長勢有正面之影響。 ❹表五杏菜種子包埋之生長狀態統計 _ -- ---對照、，组___ΒΜ種子包埋組 株南（公分） 24.4 士 1.30 27±1.41 葉片數（片） 7.1±0.24 7_6 士 0.21 根長（公分） 32.4±1.20 26.4±2.01 植株乾重（克） 26±2.03 35±2.31 實施例四田間應用情形 試驗方法： 取實施例二所得之ΒΜ包埋種子(辭)進行試驗^試驗方法如下：一分 地分作三種處理，每⑨料行種植，肥培_台肥四十三航學肥料作追 肥。三種處理分別為對照組(未包埋），ΒΜ種子包埋組及他牌枯草桿菌 (Backus subtiHs)，以600倍稀釋後澆灌作物。 調查方法： 作物定植後調查生長情形及收穫產量。 表六茄子種子包埋之收穫量統計 15 201010598 對照組 枯草桿菌 ΒΜ種子包埋組 第一次採收（公斤）24.3 19.6 36 第二次採收（公斤）11.6 11.4 15.8 總計（公斤） 35.9 31 51.8 結果： 如表六所示，茄子之bm種子包埋組之產量比對照組及他牌栝草桿菌 鬲。故，以蕈狀芽孢桿菌，BM)包埋植物種子，可促進植 株生長’植株本身較為強壯，抵抗逆境能力較強，因此收穫量較高。 〇 本發明所提供之一種促進植物生長及抵抗病源菌感染之種子包埋方法 及其產物’與前述引證案及其他習用技術相互比較時，更具有下列之優點： 本發明所提供之一種促進植物生長及抵抗病源菌感染之種子包埋方法 及其產物’其中經蕈狀芽孢桿菌包埋均勻之植物種子，種子發芽後立即受 到感染’接種率（菌數可達lxl〇9cfli/粒）較田間直接施用為高。 本發明知:供之一種促進植物生長及抵抗病源菌感染之種子包埋方法及 其產物’可於種子萌芽之初即可促進植物生長及抵抗病源菌感染β 參 本發明提供之一種促進植物生長及抵抗病源菌感染之種子包埋方法及 其產物，可自動化大量生產優良生長力之種子，免去種子浸入及乾燥等繁 瑣之程序。 上列詳細說明係針對本發明之一可行實施例之具體說明，惟該實施例 並非肖本發明之專繼圍，凡未麟本發明技藝精神所為之等效實 施或變更，例如：添加劑、植物種子等實施例，均應包含於本案之專利範 圍中。 16 201010598 綜上所述，本案不但在崎上確屬綱，並能較㈣物Μ進上述多 項功效’應已充分符合新馳及進步性之法定發明專觀件，纽法提出 申請’懇請貴局核准本件發明專利申請案，以勵發明，至感德便 【圖式簡單說明】 無 【主要元件符號說明】 無 參Embedding group 24.3+1.47 7·6 ±0.58 32.8±2.14 24±1.〇4 Control plant height (cm) 2611.55 ® number of leaves (sheet) 7.5±0.87 Root length (cm) 25.2±1.44 Plant dry weight (g 20+1.69 3. The number of rapeseed is higher than that of the control ginseng. The growth of the φ vegetable seed is embedded in the bear----the control group φ plant height (cm) 35_8±2.13 number of leaves (piece) 5.6 ± 0.25 root length (cm) 30.4±1.35 (g) 3011.01 Embedding group 37.3+1.54 5.3+0.36 25.5±1.24 28±1.60 4. As shown in Table 4, the BM seed embedding group has little difference in root length. Other growth parameters showed that the growth potential of the spinach was better after embedding the seeds with BM. Table 4: Growth status of seedlings embedded in water spinach 201010598 Plant height of BM seed embedding group in control group (cm) 66.7±2.17 71.5±2.22 Number of leaves (sheet) 32±2.03 34.7±2.57 Root length (cm) 30.9±1.39 28.2± 1. 〇1 Plant dry weight (g) 38±1.32 4511.63 5· Apricot vegetables as shown in Table 5, except for the root length of the apricot vegetable seed-embedded group, the other growth parameters are better than the control group. Therefore, it is confirmed that embedding the seeds with ΒΜ has a positive influence on the growth potential of apricot.生长 Table five apricot seed embedding growth status statistics _ -- --- control,, group ___ ΒΜ seed embedding group plant south (cm) 24.4 ± 1.30 27 ± 1.41 number of leaves (piece) 7.1 ± 0.24 7_6 ± 0.21 Length (cm) 32.4±1.20 26.4±2.01 Plant dry weight (g) 26±2.03 35±2.31 Example 4 Field application scenario Test method: Take the ΒΜ ΒΜ 种子 种子 ( ( ( ( ^ ^ ^ ^ ^ ^ ^ ^ : Divided into three treatments, each 9 rows of planting, fat culture _ Taiwan fertilizer forty-three aviation fertilizer for topdressing. The three treatments were control group (not embedded), alfalfa seed embedding group and other brand of Backus subtiHs, which were diluted with 600 times and then watered the crop. Survey method: After crop planting, investigate the growth situation and harvest yield. Table 6 Harvest statistics of eggplant seed embedding 15 201010598 The first harvest of the B. subtilis seed embedding group (kg) 24.3 19.6 36 The second harvest (kg) 11.6 11.4 15.8 Total (kg) 35.9 31 51.8 Results: As shown in Table 6, the yield of the bm seed-embedded group of eggplant was higher than that of the control group and his brand. Therefore, embedding plant seeds with Bacillus licheniformis (BM) can promote plant growth. The plants themselves are stronger and stronger against adversity, so the yield is higher. The seed embedding method and the product thereof for promoting plant growth and resistance to pathogenic bacteria provided by the present invention have the following advantages when compared with the aforementioned cited documents and other conventional techniques: The present invention provides a promoting plant Seed embedding method for growing and resisting pathogenic bacteria infection and its products 'In which plant seeds are uniformly embedded by Bacillus licheniformis, the seeds are infected immediately after germination, 'inoculation rate (number of bacteria up to lxl 〇 9cfli / grain) compared to the field Direct application is high. The invention provides a seed embedding method and a product thereof for promoting plant growth and resistance to pathogenic bacteria infection, which can promote plant growth and resist infection of pathogenic bacteria at the beginning of seed germination. And the seed embedding method and product thereof against the infection of the pathogenic bacteria can automatically mass-produce the seeds with excellent growth force, and avoid the cumbersome procedures such as seed immersion and drying. The detailed description above is a detailed description of one of the possible embodiments of the present invention, but the embodiment is not intended to be an equivalent of the invention, and equivalents, such as additives, plants, Examples of seeds and the like should be included in the scope of the patent in this case. 16 201010598 In summary, this case is not only in Saki, but also can be used to more than the above-mentioned multiple functions of the above-mentioned multiple functions. The bureau approves the invention patent application, in order to invent the invention, to the sense of virtue [simplified description of the schema] no [main component symbol description] no reference
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