TWI373304B - Method for selecting flooding-resistant papaya plants - Google Patents

Description

1373304 t VI. Description of the invention: [Technical field to which the invention pertains] Anaerobic adversity adjustment The present invention is a method for breeding (iv) flood-tolerant papaya varieties having flooding resistance by treating papaya seeds with alcohol, Give seeds [prior art] Papaya (6) Xinqing-L unveiled _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ It will rot, and then the whole plant will wither and die. Therefore, in the cultivation management, the county lion water reflects the soil cut f loam, the mechanical axis is made high wow and drainage ditch, and the water is dredged at any time to prevent root rot (Wang Denan. Cong. Papaya. Taiwan farmer's view of farming (2). Corporation Fengnian Society. pp i_6). Taiwan is due to the fact that it is located in Europe and Asia. She is expected to reduce the amount of earth and stone in the affected areas by the strong winds and heavy rains that were carried by the winds. In addition, in the agricultural sector, in addition to blowing down crops and farmland houses, the heavy rains carried by them have made the crops suffer from flooding and adverse weather. If the breeding method is selected, the _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Traditionally, there are also varieties that have the ability to withstand flooding from wild. Papaya. But because of the fruit flavor of the wild melon and the sweetness of the oral sweets, it is still hoped that Wei has fruit products f_breeding. Renovation, without getting the characteristics of the original variety, 'get the submerged papaya improved strain, but there have been no successful cases. 1373304 t Flooding is an important environmental slavery that affects the growth of art. Plants cause stomata to inhibit photosynthesis, causing the leaves to yellow. In addition, the furnace is in an anaerobic state, and the lion's aerobic respiration _, the spurt of the abundance of oxygen, produces a large amount of paving, and the material B is damaged by the cell, and the blast line is destroyed, resulting in plant death. However, the flood-tolerant plants have a high alcohol metabolism capacity, avoiding excessive alcohol accumulation in the body and causing toxic effects (Barta, A. B. 7: 187·191, 198 仏 anaerobic pretreatment can improve many plants in ^ T^^(Blokhina, OB, E.Vir〇, ainen ^ KV Fagerstedt ^

Bot. 91: 179-194, 2003) 〇 ’

During flooding, a large amount of root metabolism is most important for plant survival 'because this reaction provides more energy to maintain root metabolism (4) coffee, B. and M. C Drew, pp. 4叩8 in: τ. τ. K〇zl〇wski Μ ), flooding and plant growth. Aeademie Presslne., NewY, 1984). It is classified as the final product of anaerobic respiration of plants. Therefore, the first invention of the present invention can apply alcohol treatment to seeds, giving seeds pure adversity_lower seams, improving the water-resistance ability of the new-born plants, and seeking the ability of plants to withstand flooding. Physiological screening indicators to accelerate the selection of flood-tolerant varieties. The current application of alcohol treatment involves inhibiting fruit ripening (Saltvdt, M Ε· and A. R. Sharaf, J. dm. Soc. // 〇外) 17: 793-798, 1992.), increasing seed germination rate (Miyoshi, K. and T. Sato, et al., G. 79: 391-395, 1997), Improving plant cold tolerance (Chin, J. Η. and DB Goldstein, 196: 684-685, 1977, Chin, JH) And DB, Mo/. P/wr is co/· 19: 425-431. 1981), resistant to pests and diseases (Margosan, DA, "Γ. L. Smilanick, GF Simmons and DJ Henson, P/awiZ% 81: 1405- 1409, 1997), etc., is not applied in the 1373304 • < - high plant flooding capacity. SUMMARY OF THE INVENTION The present invention is capable of producing an alcohol by anaerobic fermentation under the condition of drowning, so that the present invention utilizes alcohol to treat papaya (a fruit tree that is extremely resistant to flooding). It is suitable for silk-to-oxygen stress, and the ability of the plant to absorb water is enhanced. In one aspect, the present invention provides a green for breeding a flooded papaya plant having flooding resistance, the method comprising: (4) papaya seeds are infiltrated with water and then placed in a 〇·25 ml/4L Treat to 2 ml/4L of anhydrous alcohol vapor at % C for 1 to 3 days, sow the alcohol-treated seeds to germination; select the papaya seedlings grown to 6-8 leaves for flooding for 2 days. Drainage is provided; and plants that grow well after drainage are selected as flood-tolerant papaya plants. In the specific aspect of the present invention, the alcohol vapor concentration used is preferably 0.25 ml / 4 L to i ml / 4 L 'better _ 〇 5 _L to 1 匕. In the case of the present invention, the treatment time of the alcohol vapor is preferably 2 to 3 days, and the treatment period of 3 days is the best. In another aspect, the present invention provides a method for improving the flooding resistance of papaya plants after germination, which comprises applying papaya seeds to a concentration of 〇25 ml/4 L to 2 ml/4 L before germination. Alcohol vapor treatment for 1 to 3 days. Other features and advantages of the present invention will be further exemplified and illustrated in the following embodiments, which are merely illustrative and not intended to limit the scope of the invention. EXAMPLES The following examples are taken from 'Tai Nong No. 2', 'Risheng', 'Ex〇tica', and papaya seeds of different varieties, and treated with different concentrations of alcohol. The method is as follows: Before processing, firstly, Tainong No. 2 ', 'Rising' and 'Exotica, the papaya seeds were placed in a 25 〇ml beaker for 24 hours under running tap water. The infiltrated papaya seeds were placed in a petri dish for alcohol treatment. The Petri dish was placed in Advantec N〇. No. 1 diameter 9 cm Luzhizhi paper. The seeds were placed on filter paper and the filter paper was wetted with Sml pure water. Place the culture dish in a 4L sucking tank, take 〇25m dip (four), lm 2ml pure alcohol (anhydrous alcohol) in a beaker, 'the other group is the control group without adding alcohol, then put the beaker in the breath The alcohol is volatilized in the cylinder, and the sucking cylinder is sealed immediately after the alcohol is processed (three dishes per treatment, and two seeds per seeding). Put the closed breathing cylinder in the Deng C incubator' during the breathing of the rainbow seal!, hour, day, 2 days and 3 days. After the seed is released, the seedlings are treated with #木绿水水, flooding method - in order to use 8 liters of plastic pots, the plants of different varieties are placed in the basin, and the water is given, • the water surface height is flooded over the soil surface. The flooding time is 2 days, and stones are placed in each of the three bowls to avoid plant tilt. After draining, observe the flooding tolerance of the papaya plant and analyze the relevant physiological changes. The following examples were given for 'Tai Nong No. 2, seed spot _ strains were 3 replicates, treatment group was CK (no alcohol treatment control group), Id (breathing cylinder sealing days) 〇 25 (ml/4L) ), ld 〇5 (mi/4L), w I (ml/4L), Id 2 (ml/4L), 2d G.25 (ml/4L), 2d 0.5 (mI/4L), 2d 1 (ml/ 4L), 3d 0.25 (ml / 4L), 3d 0.5 (mI / 4L), 3d l (ml / 4L), a total of U treatment. The alcohol treatment group of 'Rising' seeds was 0, 2d (breathing cylinder sealing days) 1373304 k 0.25 (ml/4L), 2d0.5 (ml/4L), 2d l (ml/4L), a total of 4 species deal with. Example 1: Treatment of papaya seeds with different alcohol concentrations and their effects on seed germination rate Seed germination was observed and recorded until the seeds were no longer germinated, and the average germination days and total germination rates were calculated. The average number of germination days is t = S (t · η) / Σ η, where t: days after bed setting; n: number of germinations on day t after bed setting; Σ η: total number of germinations during germination test. Total germination rate = (number of germinated seeds / number of seeds placed after the test) χ 1〇〇% After 1 day of alcohol treatment of 'Tai Nong 2' papaya seeds, the seeds of the control group germinated faster. The germination was slower, but the total germination rate of the seeds treated with 〇.25mi, 〇5ml, lmi alcohol was similar to that of the control group, and the germination rate of 2ml treatment was low. After 2 days of alcohol treatment, the 'low concentration aki _.25, a5m_# test seeds There was no significant difference in germination rate. The total germination rate of G.25 and 〇.5ml alcohol treated seeds was even higher than that of the control seeds. However, the seed germination of high concentration treatment (lml) was still slow, and the germination rate was also low. There is no germination (the result is magical). 'Nippon, papaya seed 2 treatment 1 or 2 days, G.25, G.5, lml seed germination situation similar to the Lin Zhao group, the germination began 7 days after the seed, the total germination rate was not significantly different from the control group, However, if the seed has a low or no germination rate. At 3 days of treatment, the control group had no significant results: Γ二=Parts ° ° 25, °. 5 ml treatment and control group 2 (see Table D ° *Ex〇tica, ^^^ Most low-concentration alcohol relied on Cards and privately lose M f to see 'day time, (four) 赖 group and money difference, and fine treatment 2 •,. The treatment germination rate will be higher than the control group, too high alcohol concentration fine treatment seed material low or no Budding (Table 1).

The volume of alcohol evaporating in a 4L container is treated for i to 3 days. The same letter of the average value of each paste in the paste indicates that they are 5°/ when compared with the (10) (10) secret (four) alcohol test. Nothing on the level < no germination. ',,,·,different. 504 A combination of the above test results, found that 〇2M, 〇5mi de concentration alcohol treatment, high papaya seed hair, but no significant effect on the germination rate; relatively, excessive j alcohol concentration treatment will reduce the germination rate and germination of the health It is not even louder to inhibit seed germination. Agriculture - said ',, 曰陛, or, Ex〇tica, the variety of papaya seeds by alcohol treatment =, the rate of material (four) decreased or not issued ^ low concentration bribe treatment can promote seed hair #细柯胄b7C mesh for alcohol Processing helps the seed break sleep. Appropriate alcohol concentration can increase the germination rate, mainly because of the permeability of Qi (4), affecting the role of membrane lipids or specific sites on the sputum, ethanol, C - these fat-soluble substances, can quickly affect the seed material, or It is related to the germination and metabolism of stimulating seeds. Lai can promote the absorption of oxygen and accelerate the material, and increase the concentration of fructose_2,6·diquaternary _uCt〇Se2'6-bi_spoate) to stimulate the glycolytic effect of the seeds of Hume. Example 2: Seeds _ fine New Zealand's Mumu New strain in the drowning before and after the change of seeds without wine secrets on the ship, 'ExGtiea, H, and 4th "variety papaya plants after flooding for 2 days" The leaves will obviously sag yellow, the plants will be skewed, and the drainage (four) pieces will be severely withered or even die. In the treatment part, after the seeds are treated with different concentrations of alcohol and different sealing days, the subsequent plant flooding performance will be different. The flooding changes vary among varieties. In the case of Έχ—, for plants, ld〇.25ml alcohol treated plants drooped significantly on the 2nd day after flooding, and leaves became yellow after drainage. But the symptoms of 2d 0.5ml were mild, and after 6 days of drainage, Id 〇 25 plus

The plants of Idhni are severely withered, and the leaves of plants treated at other concentrations are still drooping. The plants treated with 'Tai Nong No. 2' had no significant change in the first day of flooding. On the second day of flooding, except for the LD 0 and ld (4) treated plants, the other plants 1373304 I » remained in normal condition (Fig. 1). On the 4th and 6th day of drainage, the leaves of ld G 25mi, id 0.5m, Id lml alcohol treated plants were severely drooping, and the dish was 5mh3d - there was no significant change after flooding and drainage (Fig. 2). 'Sunday, the alcohol-treated plants are similar to 'Tainong No. 2' and there is no obvious change after 1 day of flooding. After 2 days of drainage, id 〇 25mi, id 〇.5, ld lml alcohol treated plants sag Severe (Fig. 3), and on the 4th day of drainage, the leaves of Id 0.25ml and Id 0.5ml had their leaves _ withered, and the drainage was more serious after 6 days of drainage, and the 3d 0.25ml alcohol treated plants were also drained 6 After the beginning of the day, the fresh 'Il lml, 3d G.5m Bu 3d 1ml alcohol treatment, the plant growth is better (Figure 4). Papaya plants are extremely resistant to flooding due to their fleshy root characteristics. After 24 to 48 hours of root immersion, they will start to cause death. After the 48-hour flooding treatment, the papaya plant drastically yellowed and drooped, eventually causing the plants to wither and die. However, different varieties of papaya have different ability to withstand flooding. The performance of 'Exotica' papaya flooding is earlier and more serious, while 'Tainong No. 2' belongs to moderate flooding adaptive group. It is known from the results of this experiment that the pre-treatment of papaya seeds can delay the flooding symptoms or increase the survival of the plants, indicating that the seed anaerobic pre-treatment can help the plants to withstand flooding stress. Example 3: Chlorophyll content and fluorescence change of papaya plants treated with alcohol vapor before and after flooding The chlorophyll content of papaya plants after flooding was significantly reduced, which resulted in yellowing of papaya leaves after flooding in this experiment. One of the reasons is that in this example, the leaf chlorophyll content was determined by SpAD5〇2 (Chlorophyll meter) Minota to observe the effect of alcohol treatment on the leaf chlorophyll content of papaya plants before and after flooding.

The results showed that the 'Exotica', 'Tai Nong 2, and 'Supreme', papaya seeds without alcohol treatment showed significant chlorophyll content in the leaves before and after flooding, 'Ex〇tica, plant in flooding 2 The chlorophyll was significantly reduced, 'Tai Nong No. 2, and, ascending, the plants were significantly reduced after drainage (see Table 2 to Table 2_3 below). Treatment of different varieties of papaya seeds with alcohol 'In addition to a few lower-contaminated plants, the chlorophyll decreased after chlorophyll reduction. Most of the plants treated with noodles did not change significantly before flooding (4). Compared with the postal treatment of 5, the 'ExGtlea' bribe treatment and the control group had no difference in chlorophyll content before flooding and the first day of flooding, but on the 2nd day of flooding and 4-6 days after drainage = 3,d 25.25ml plants have the highest chlorophyll content (see Table. 'Tai Nong No. 2, Alcohol treatment, ', the chlorophyll content of plants before flooding is lower than that of the control group, and after 5% of days after drainage, the chlorophyll of quercetin treated plants The content will be higher than the control group (Table Μ). 'Yilu, plant chlorophyll = fine treatment and control plants before flooding, 2 days of flooding and 2 days of drainage, there is no significant difference in plant t' but in drainage After 4 days and 6 days, 3d lml alcohol treatment was higher for flooding chlorophyll (Table 2_3). These results show that alcohol treatment can improve the tolerance of some plants to H and its chlorophyll content is not affected by flooding stress. Treating 'Exotica' papaya plants before and after flooding

BFX — Flw i^l5auA 44.73abA d6.65aA _ 45,83aA~ i〇-〇7aA _ 3M3bATM j6-70aA_ 45.65aA j3.83aA_ 43.80abA i!48aA 44.43abA 44.60abA 47.15abA 48.95aA Chlorophyll (value) D2 ' D4 D6

40.05bB

40.10bAB

39.80bA

46.10abA

48.13aA

45.48abA

38.98bB

39.45bAB

39.90bA

47.05aA

41.7bA

47.90aA

38.93abB

35.65bB

39.17abA

40.48abA

40.23abA

45.50aA ll 1373304 4 z Evaporate the volume of alcohol in a 4L container for 1 to 3 days. yCK=no alcohol treatment. XBF=Papaya plant before flooding WF1=Papaya plant was flooded for 1 day. VD2=Papaya plants were flooded for 2 days after flooding for 2 days. . The multiple range test listed in the ^ (lowercase) and f (column) average values is statistically compared at the level of 5%. (10) Coffee • ^^ Fine steam treatment 'Tainong No. 2, Papaya plant Before and after flooding

Treatment zn·recording, (day, ml/4L) BFX Flw F2 D2V D4 D6 CKy 50.03auA 47.33aAB 43.43aAB 41.20bAB 40.65bAB 37.25bB Id 0.25 40.90bA 43.05aA 44.18aA 39.33bA 40.28bA 43.15abA Id 0.5 46.65abA 41.90aA 43.70aA 42.33bA 42.35abA 40.43abA ldl 43.68abA 43.60aA 44.98aA 45.40abA 43.73abA 38.10abB 2d 0.25 43.90abA 47.88aA 46.15aA 46.83abA 44.50aA 45.85abA 2d 0.5 45.15abA 43.35aA 45.48aA 47.50aA 45.70aA 45.40 abA 2d 1 44.47abA 44.70aA 46.23aA 45.43abA 43.07abA 44.17abA 3d 0.25 42.23bA 43.90aA 45.25aA 47.25aA 44.00aA 46.38aA 3d 0.5 45.35abA 44.93aA 46.30aA 47.93aA 45.70aA 43.45abA 3dl 46.87abA 46.40aA 46.73aA 47.50aA 47.97aA 47.70aA 12 1373304 • Table 2-3. Chlorophyll changes in seeds treated with alcohol vapor. Soaring 'Papaya' plants before and after flooding

D6_ 39.1〇bB 39.90bB 38.98bB 42.93abA 39.53bC 42.45abA 44.93aA Chlorophyll fluorescence ignoring and non-destructive failure method, we can know the son-in-law efficiency of the PS Π system in photosynthesis systems, and the system in the case of adverse or adverse conditions. The medium protein • repair situation is reflected in the fluorescence parameters and can be used to screen indicators of crop stress tolerance and assessment of plant recovery after stress (Baker, NR and E., J Exp Bot • 1607-1621.). The analysis method is briefly described below. First, the leaves were subjected to dark-adapted treatment using leaf darkness, and the (d) Portable DtoorphyU fluGn meter, ΜΜ_ρΑΜ, _ζ, Germany) was turned on for chlorophyll fluorescence measurement. The connected probe Xiang special gamma gamma Fiberoptic 2〇10-F) provides the measured light and saturated pulsed light instantaneously, and is calculated by the following formula to obtain PS Π activity (expressed as Fv/Fm). °

Fx-Fm-Fo' is expressed in FWFm (Fq is the minimum amount of fluorescence emission, and Fm is the maximum amount of fluorescence emission). 13 1373304 The results show that 'papwood seeds without alcohol treatment, whether in the &, sa Nong 1' or 'day ft' plants, their Fv/Fm fluorescence values will be obvious during the drainage period after flooding Reduce the number of seeds treated with alcohol from 0.80-0.82 to 0.66-0.68 (see Table 3·ι to Table 3 3 below), except for ld G.25m b · 25m b 3d G 25m 丨 low concentration alcohol treatment After the fall of the fluorescing, the rest of the sputum treatment = chlorophyll camping light did not change significantly; if the same treatment time, there is no difference in the presence or absence of alcohol treatment in the flooding 2 days, but after drainage, large Some alcoholic slaves were still _higher, and only had a decrease in the Wei value of the alcohol treated plants. Papaya decreased after flooding and drainage, and it was known that it was damaged by flooding, affecting photosynthesis and reacting to the performance of parameters. However, the Fv^pm values of the alcohol-treated plants before and after flooding did not change significantly or higher than that of the control group, indicating that they were not damaged by the stress or the plants recovered well after drainage.士3_y Seeds treated with alcohol vapor, ‘Ex〇tica’ papaya, whose leaves are green and silky.

Treatment z Fv/Fm (day, ml/4L) BFX Flw F2 D2V D4 D6 CKy 0.82auA 0.79aA 0.75bAB 0.73bB 0.69bC 0.66bC Id 0.25 0.81aA 0.79aA 〇.78abA 0.72bB 0.74abB 0.74abB ldl 0.83aA 0.81 aA 0.83aA 0.80aA 0.80aA 0.80aA 2d 0.25 0.80aAB 0.81aAB 〇.82aA 0.80aAB 0.77aB 0.79aAB 2d 0.5 0.81aA 0.80aA 0.80aA 0.78abA 〇.74abA 0.79aA 3d 0.25 0.81aA 0.80aAB 0.81aAB 0.79abAB 0.77 aB 0.77abB yCK=no alcohol treatment. XBF=Papaya plant before flooding WF1=Papaya plant was flooded for 1 day. VD2=Papaya plants are flooded for 2 days after flooding for 2 days. u The letters listed in each column (lower case) and the average value in each column (capital) are statistically compared with Duncan's multiple range test at 5%. Make a distinction on the level. r 14 1373304 t . Table 3-2. Chlorophyll fluorescence changes of 'Tai Nong No. 2' papaya seeds treated with alcohol vapor before and after flooding.

Treatment z (day, ml/4L) Fv/Fm BFX Flw F2 D2V D4 D6 CKy 0.80auA 0.81aA 0.77b AB 0.71cBC 0.73bBC 0.68cC Id 0.25 0.80aAB 0.82aA 0.8 lab AB 0.72bcBC 0.70bC 0.70bcC Id 0.5 0.81 aA 0.82aA 0.81abA 0.78abA 0.81aA 0.77abA Id 1 0.81aA 0.82aA 0.81abA 0.77abA 0.80aA 0.79abA 2d 0.25 0.80aA 0.81aA 0.81abA 0.81aA 0.78abA 0.72bcB 2d 0.5 0.81aA 0.82aA 0.80abA 0.80aA 0.80 aA 0.76abA 2d 1 0.82aAB 0.82aAB 0.82aAB 0.81aB 0.81aB 0.83aA 3d 0.25 0.83aA 0.82aAB 0.82aAB 0.79aBC 0.79abBC 0.77abC 3d 0.5 0.82aA 0.82aA 0.80abA 0.80aA 0.81aA 0.77abA 3d 1 0.82aA 0.82 aA 0.82aA 0.82aA 0.81aA 0.82aA Table 3-3. Changes in chlorophyll fluorescence of leaves of 'Zisheng' papaya seeds treated with alcohol vapor before and after flooding.

Treatment z (day, ml/4L) Fv/Fm BFX Flw F2 D2V D4 D6 CKy 0.81auA 0.79bAB 0.77bABC 0.74bABC 0.71bBC 0.68bC Id 0.25 0.82aA 0.81abAB 0.79abAB 0.81aAB 0.73bB 0.74abB Id 0.5 0.83aA 0.82 aA 0.79abA 0.81aA 0.77aAB 0.70bB Id 1 0.81aA 0.78bAB 0.81aA 0.82aA 0.77aAB 0.73abB 2d 0.25 0.82aA 0.80abAB 0.81aAB 0.78abB t - 2d 0.5 0.83aA 0.80abA 0.82aA 0.78abA - - 2dl 0.81aA 0.80abA 0.82aA 0.80aA - - 3d 0.25 0.83aA 0.82aAB 0.82aA 0.81aAB 0.77aB 0.72abC 3d 0.5 0.84aA 0.81abA 0.82aA 0.80aAB 0.73bC 0.78abB 3dl 0.81aA 0.81abA 0.82aA 0.82aA 0.76abA 0.80aA 15 Package IV. Changes in mineral π-seeds and/or oxygen before and after flooding of papaya plants with seeds at the bribes of alcohol and alcohol will inhibit aerobic respiration and reduce the absorption of inorganic nutrients (Nilsen'ET water, MA〇 Crcutt'pp 362_400, John Wiley and S〇ns, Inc., 1996). Flooded. Roots. P oxygen partial pressure will be reduced, reducing cellular respiration, Zhonghe quotient (four) fine town X children eight, release C 〇 2 / absorption 〇 2) will increase a lot. The new root and the apex have a higher pro-pressure only because the respiratory rate is high and there is no air chamber between the birds. Therefore, after flooding, the partial pressure of the milk will increase rapidly. This is related to the absorption of nutrients. The environment was characterized by the first month and the first symptom was the yellowing of the leaves. In the years of ι〇月4, No. 2, before plant flooding), 6th ('Tainong No. 2, two days of plant flooding), 8th=Nong'e 2' plant drainage for two days) and fine years 12 On the 5th of the month ('plant flooding 2,7曰('曰©, plant flooding for two days), 9曰('曰?, plant drainage for two days) for picking. Papaya plants will be attached to the dust by tap water Cleaned, then cleaned with i%ira fans to wash the silk with deionized tree for 3 times. _ Moisture is packed in a kraft paper bag. As for the ventilated oven, first kill the cyanine and then adjust the temperature to 7〇C, until the sample·. The dried branches are ground into a powder by a mill, and the dried leaves and the roots are weighed into a crucible for elemental analysis.

After the flooding of the papaya plants in this experiment, the nitrogen concentration in the leaves and roots may increase or not change significantly. However, if the same treatment time is compared, before the second flooding, the non-alcoholized papaya seeds are treated with the leaf axillary Z-treated leaves, and it is speculated that the alcohol pretreatment makes the leaf concentration in the control group. High, _ after flooding or drainage, there is no significant change in the number of plants in many simple treatments. The ability to display nitrogen is not affected (see Table ^ 3 16 丄 373304 * . dd y changes, regardless of the shirt After contact with spermatogenic treatment, the phosphorus concentration in the roots and roots increased or did not change significantly after flooding and drainage. The posterior leaves and papaya plants were expressed in the 3dl before flooding, where zx, w, y, v and u were represented; Before changing water:

2.76euC

2,73eB

3.83dB

3.87dB

4.46bA

3.96cdA

4.20bcA

3.65dB

3.80dA

4.37bAB 4.93aA

f flakes 3.60abB 3.41bA 3.61abC 4.23aA 4.20aA 3.81abA 4.29aA 4.12abB 3.70abA 4.26aB

3.73abC 4.39bcA

3.67eA

4.25cdA

4.27cdA

4.35bcA

3,78eA

j!4dA

1^4cdA 4.55aA —

4.5〇abB

BF 2.56bcdA 2.01dA 2.99abB _2.78abcC 3.39aB _2.59bcdB 2.53bcdR 2.17cdC _2.36bcdB 2.60bcdB 2.77abcA a jp

2.34cA _2.44cA _2.81bcB _3.01aB^ _3.22abC 3.60aA A 2.40aA — _3.1SabA 3.3SabA

239cB

Drainage 3.45bcA 2.66dA 3.69abA 3.98aA 3.61bA 2.71sB ^33bcA 2.60dB _3.19dA j.49dB Leaf ί and root ώΐϊϋ treatment of ‘日陆’ 瓜瓜丝 before and after flooding

17 1373304 z The volume of alcohol evaporated in a 4L container for 2 days. yBF = papaya plant before flooding x flooding = papaya plant was flooded for 2 days. I Drainage = Papaya plants were flooded for 2 days after flooding for 2 days. The letters listed after the average value of 内, 内 (lowercase) and each row (capital) are closely distinguished at the level of 5% when compared statistically by the dirty multiple test. The unloading concentration side is financial, and some treatments will increase, while others will decrease or not change significantly. In the change part of 'Tai Nong 2' papaya plant, the results showed that Id 0.25ml, id 0.5m 2d 0.25ml planting_ concentration will decrease after drainage, while id lmh ld 2ml, 3d 〇.5ηύ, 3d lml will Increased (Table 4·3); however, there was no significant change during the flooding during the day, and the 〇, lml treatment (Table 4_ ::

3 related, 'Tai Nong 2, and 'Day M, # papaya plant roots, all view = 酉 酉 之 之 之 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 钾 ( ( ( The reason for the change before or after the water is not large or increased. Α 4·4) 'And 1373304 • · The seed of the table is treated with alcohol vapor, 'Tainong No. 2, # papaya, the plant is drained and the roots are unloaded. Variety.

The meanings represented by 2, \, evaluation, 丫1 and 11 are the same as those in Table 3_1 above. Table 4-4. The seeds were treated with alcohol vapor, and the changes of potassium concentration in leaves and roots of papaya plants before and after flooding were observed. .

Where z, y, x, w&v represent the same meaning as Table 4_2 above.

- Κ (%ΌΨ) Treatment z Slices (4) 15 (ml/4L) BFy Flooding x Draining w BF Flooding drainage CK 3.37bcvA 4.05aA 3.47aA 3.86bA 4,0QaA *2 04ΚΔ 0.25 2.91cB 3.61aA 3.60aA 3.91bA 3.19cB Λ 97〇A 0.5 3.91aA 3.48aA 3.53aA 3.84bAR 3.4%i>R 1 3.68abA 3.64aA 4.14aA 4.53aA _3-75abB 4.33aA As for other persuasive elements, the 番_精处理的papaya seeds The leaves and roots of the plants will be reduced after the drainage, and the soil moisture will be reduced. However, the roots of the plants treated with alcohol 19 13373304 will be reduced after the drainage, but some of the leaves in the leaves will be fed with magnesium. The concentration will increase or not change. In the change of root steel concentration, the root concentration of the 'Tai Nong 2' control group will decrease from 18 lions to U PPm when flooded, and will increase to 13 after the transfer, and its concentration after drainage. It is obviously higher than the rider. _ fine treatment is the second day of flooding, the copper concentration in the root will change or increase, and after the drainage, the Qing will decrease, · '峨, the root is submerged' The copper concentration will increase 'and decrease after draining, but the W-alcohol treated plant will remain higher after water (Table 4_5). ί and 4 部 种 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1

Example 5: Changes in respiration rate and ethylene release rate of papaya plants treated with alcohol vapor before and after flooding were sampled using a drill having a diameter of 1.1 cm, and 5 pieces of papaya leaf tissue discs were sampled and weighed. In a 25 ml flask, 5 ml of deionized water was added, and after sealing for one hour with a serum plug, '1 ml of gas in a triangular flask was extracted with a plastic syringe, and the ethylene release rate was measured by gas chromatograph (Shimadzu Model GC-8A). ' 20 丄: Early position with μΐ C2H4/kg.hr silk red material oxidized age (four) Yangcai leg spider UN〇R61_ fixed rate, single fairy (five) c added. ,

The plants with papaya seeds that have not been treated with appropriate fineness will have a significantly lower respiration rate after flooding and drainage. The changes in leaf respiration rate of the seeds treated with alcohol are inconsistent, %Nong 2, ld a25ml, 3d A5mi treatment (4) The rate of suction will decrease after flooding and drainage, but the leaves treated with other alcohol concentrations will have a tendency to increase beer suction rate (Figure 5). In the ‘Sunday, the part of the alcohol-treated plant, the leaf respiration rate changed after flooding and rowing, and the result was not, but there was no statistical change in the duck. Flooding can cause the respiration of papaya leaves to decrease, mainly because the flooding stress will increase the amount of leaf ’3, which in turn promotes the stomata of the leaves, making the leaf: oxidized carbon disk oxygen exchange significantly controlled. In the part of the root respiration rate change, except for the 'Tai Nong 2 W 2d a5ml and ' a5ml, lml alcohol treated plant roots after flooding, the respiratory rate ''', '·, 贞 change' other treatment and control The respiratory rate of the roots of the group was significantly reduced (Fig. 6, Table M). The water respiration rate of papaya roots was reduced by flooding treatment. It is speculated that the silencing b can be related to the inhibition of root metabolic activity by hypoxia stress. A day (4) before and after the tank is flooded

21 1373304 • , In addition, plants induce ethylene to produce M under flooding conditions.

-·细.μ 97:584_588, 1972.), in this experiment, the papaya plant will increase the amount of ethylene in the leaves by 2-4 times after flooding, but the amount of ethylene produced by seeds and alcohol-treated plants will be ·Below the Lai group (see Figure 7 and Table 5·2), the Ethylene part of the root is 'no difference in flooding and drainage before and after drainage. After 2 days of drainage, the alcohol content of the 'Rising' plant roots is lower than that of the control group. Low (Table 5_2). The increase of leaf ethylene is mainly due to the fact that the roots of plants are under the Wei, and it is expected that ACC will be synthesized in large quantities. The oxygen is required for the formation of ethylene, but the roots of plants are not able to produce ethylene under flooding and low oxygen, but a large amount of ACC will accumulate. ACC will be transported to the ground through the xylem to produce a large amount of ethylene, w κ, XZ million (10), and SF & He Li Qing W. 88: 553-558, 1988.), the production of ethylene will cause the film to be biased Sexual growth caused the leaves to sag, and in this test, papaya leaves were also observed to be discharged after flooding, mainly due to the use of women. The alcohol-treated plants have lower ethylene production, and the side may be related to their ability to withstand flooding. Table 5.2. Changes in Ethylene Release Rate of Leaves and Roots of Papaya Plants Before and After Flooding by Seeds Treated with Alcohol Vapor.

The meanings represented by ,, and v are the same as those in Table 4_2 above. 22 1373304 » • Example 6: Algae dehydrogenase (ADH) and peroxidase (p〇D) of 4 papaya plants treated with alcohol vapor before and after flooding Activity and total compounds (changes in jpq content ADH is a key enzyme in the reaction of alcohol to vinegar, which catalyzes the conversion of BJ to alcohol. Its main function is to be responsible for NAD+ recirculation during anaerobic respiration, so that glycolytic energy can Continued, there have been many reports that the activity of ADH is related to the ability to withstand flooding. This experiment analyzed the change of ADH activity in roots before flooding, 2 days of flooding and 2 days of drainage. The fresh weight of plant roots was lg, and the speed of liquid nitrogen was rapid. After freezing, place in a pre-cooled mortar and add

Containing 100mMHEPES (N-(2-hydroxyethyl) piperazine-Ν'-2-ethylidene acid) pH 7.4, 0.5 mM

ZnCl2, 1 〇 p p 疏 乙醇 乙醇 乙醇 、 、 、 、 二 二 二 二 二 二 二 二 二 二 二 二 二 3 3 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 甘油 于 于 于 于 于The supernatant was centrifuged at 200 〇〇 xg for 20 minutes for activity analysis. The ADH reaction buffer is sequentially added to the spectrophotometer (cuvatte): 〇.5ml 180mM Mes-KOH buffer pH 6.4, 0.1ml 10mM DTT, 0.1ml 30mMMgCl2, 〇·1ιη12ιηΜβ·ΝΑΟΗ(ρ-nicotamine adenine Dinucleotide, also Lu original shape), then add 0.1ml of the above-mentioned rope extract, mix and tick the reaction for π seconds, and finally add 0.1 ml of 29.7 Fuji to mix evenly, immediately after spectrophotometer (spectrophotometer, Hitachi) U-2000) The change in absorbance at the initial wavelength of the reaction at a wavelength of 340 nm was measured, and the reaction rate in 〇6 〇 seconds was calculated. The unit is expressed in the stupid world. The results showed that after the seeds were not _ slaves #木绿株 in the submerged water, the roots and legs 1 mixed significantly increased (Figure 8), the seeds of the alcohol-treated plants, ADH activity will increase significantly after 2 days of flooding, Higher than the control group, it indicated that the papaya plant treated with alcohol 23 1373304 increased the adh activity in the submerged submerged water faster than the control group (Fig. 8), indicating that the seed was treated with alcohol to induce the plant to be submerged in activity earlier. . After draining, except for the increase in ADH activity of Id 0.25 ml alcohol treatment, ADH activity decreased in other alcohol-treated plants (Fig. 8). There was no significant change or decrease in μη activity in the late flooding period, mainly because the new isozyme was produced at the beginning of flooding, which provided the first line of flood-resistant defense mechanism, but the complex flood-resistant The mechanism still needs other physiological, biochemical, tissue and other adjustments in order to have better flood tolerance performance (Zhuo, 2001). Compared with the alcohol treatment concentration of the same treatment time, the root activity of the 1ml alcohol treated seed was significantly higher than that of the control group and other alcohol treatment groups before flooding and 2 days after flooding, presumably seed pretreatment, and Increased ADH activity is related to making papaya plants more resistant to flooding. Plants produce hydrogen peroxide under flooding conditions. b.,t. v

Chirkova and V. Fagerstedt, Ji^p. Ga. 52: 1179-1190, 2001.), the production of reactive oxygen species can cause great damage to cells, and the plant will initiate a series of antioxidant mechanisms in order to eliminate the accumulation of reactive oxygen species in the body. , the synthesis of many antioxidants to help the plant against the adversity. Peroxidase (POD) is an antioxidant enzyme that reduces hydrogen peroxide to water. Under the influence of antioxidant enzymes S〇D, CAT, POD, etc., it plays an important role in maintaining cell metabolism balance. The peroxidase activity was determined as follows: The leaves and roots of papaya plants were weighed and placed in an ice bath. A small amount of sea sand and 5 ml of 0.1 M dish acid buffer solution (pH 7.0) were added for grinding. After the centrifugation, g _ ' was centrifuged at 2000 C x 20 min for 20 minutes at 4 C, then filtered with Miracloth (Merck), and the supernatant was taken for activity measurement. In this experiment, the activity of p 〇D in papaya leaves without alcohol treatment decreased at 24 1373304 after flooding for 2 days, but the activity of POD increased significantly after 2 days of drainage. 'The POD activity of leaves treated with alcohol was 2 days after flooding. When the time is increased, the treatment group with 0.25ml is the most obvious. If the concentration of alcohol treatment is different at the same treatment time, the peroxidase activity of the leaves of 〇.25mi and 0ml is the highest, followed by the control group, and the treatment activity of lml is the same. Lowest (see table 6-1 below). Liu (Liu Yichang, Luo Yifeng, Chen Rongfang. Study on the effects of dry and Chengjing on the protein, peroxidase and some root traits of sweet potato leaves. Chinese Horticulture 43(2). 126 140, 1997) and other scholars found that sweet potato leaves were treated by flooding After the p〇D activity also increased significantly, the results showed that flooding stress induced papaya to initiate antioxidant defense mechanism to resist adverse conditions, while alcohol-treated seeds increased plant p〇D activity faster 'this result and Amor ( Am〇r, Υ·, Μ·Chevi〇nb and a café, the age of m-war ancestors and other scholars have similar results, and found that the large culm after hypoxic pretreatment can be hypoxic Wei Survival may increase resistance to peroxidase activity against Wei Qian, but green concentration _ Ernu Na ^ active before and after flooding bribe b, _ can _ handsome camphor activity increased time New thief drowning _, wrinkle The anti-oxidation mechanism of the paste. T is the change in activity, and the activity of the 25ml alcohol-treated plants is significantly increased after flooding (Table 6-1).

25 1373304 < * ϋ· Seeds treated with alcohol vapor ‘Sunrise, Papaya plants have changes in the activity of the gasification enzymes in the bracts and roots before and after flooding. The volume of alcohol in the 4L container of the wheat secret, processed for 2 days

0.25 ~ 53.62auB 6.29abB 20.76bcA 1 33.71bC 55.56aA 54.51aA

10.76cA

yPOD = remaining X

15.01cA Root Drainage v BF Flooded Drainage 82.95aA 37.80aA 35.96abA 30.30abA 69.60abA 25.22aB 57.03aA 29.77bB 39.8 lbcA 44.37aA 33.21bA 40.53aA 11.61cA 44.45aA 45.86abA 35.44abA Peroxidase w, v and u The representative meaning is the same as the above-mentioned Table 34. The expected compound is a secondary metabolite, which is widely present in plants. Its content can provide plant protection and can be used as an antioxidant (Takahama, U and τ 〇 11 «0 , /% ^ / > / pro 101: 845_852, 1997), this test found that after the flooding treatment of papaya, the total phenolic compounds in the leaves and roots did not change significantly before or after flooding or drainage. However, if the concentration of alcohol in the same treatment time is different, the content of total phenolic compounds in the roots before flooding and after 2 days of flooding, the lowest dose of 0.5 ml and 1 ml of alcohol treated seeds treated with the control group and 25 ml of alcohol. The total phenolic compound content of the plant is high (Table 6-2), so it is speculated that alcohol pretreatment can improve the flooding ability of papaya, which is related to the antioxidant defense mechanism. 26 1373304 I » Table 6-2· Seeds treated with alcohol vapors ‘Swelling, the change of total phenolic compounds in leaves and roots of papaya plants before and after flooding.

TPCyrue/e.FW^ treatment z-leaf root (ml/4L) BFX 1 flooding w drainage v BF flooding drainage CK 3575.93auA 3076.77aA 3052.56aA 113.20bA 46.98bA 96.29aA 0.25 3542.57aA 3835.09aA 3844.03aA 152.92bA 52.79bA 89.63aA 0.5 3755.58aA 3098.72aA 3317.85aA 19l.62abA 102.64aA 166.97aA 1 3868.54aA 3285.42aA 3804.59aA 260.50aA 116.93aA 111.39aA φ ζ The volume of alcohol evaporated in a helium vessel was treated for 2 days. yTPC=Total phenolic compounds The meanings of the remaining X, w, V and U are the same as those in Table 3-1 above. Example 7: The total free amino acid and soluble of papaya plants treated with alcohol vapor before and after flooding Changes in protein content The total free amino acid (FAA) was determined as follows, using Rosen (Rosen, H., the sinking / ^ muscles of the outside of the 67:10-15. 1957.) The p〇D filtration supernatant obtained in the above Example 6 was diluted and mixed with 0.9 ml of deionized water, and then added with 1 ml of Ninhydrin reagent (the preparation method is as follows: 5 g ninhydrin, 95 g KH2P04) , 43gKHP04, 3g fructose dissolved in 600 ml of deionized water, then quantified to il, stored in TC), and placed a marble on the test tube to prevent water from seeping, put in boiling water for 10 minutes, remove and quickly cool 'Add 5ml of color thinner (the preparation method is as follows: 2gKI03 dissolved in 600ml deionized water, then quantified to 95% with 95% alcohol), shake evenly, and absorb at 570nm by spectrophotometer (Hitachi U-2000) The value changes. 27 1373304 Standard curve was prepared with 1 mM alpha-alanine. The unit is expressed in millimolar acid/g.FW. The soluble protein (SP) was determined by the method of Lowiy et al. (1951). Take 0.1 ml of POD filtration supernatant and add 1.9 ml of deionized water to dilute and mix. Add 5 ml of reagent A (2g Na2C03, lml K2C4H406). (2% potassium tartrate), lml CuS04 (l% CuS04 · 5H20), 10ml IN NaOH, 90ml H20) evenly shaken 'after standing for 10 minutes' then add 〇.5ml reagent B (folin reagent: H20=1: 1) The oscillation was uniform, and after standing for 30 minutes, the change in absorption value at a wavelength of 660 nm was measured by a spectrophotometer (Hitachi u_2〇〇〇). The standard curve was prepared at 2525 mg/ml BSA. The unit is expressed in mg/g.FW. A hypoxic or anaerobic environment inhibits the synthesis of previously existing proteins, which in turn induce the synthesis of new proteins (ANPs). The results of this experiment showed that the seeds were treated with no alcohol and 〇.25ml alcohol treatment. The leaves of the melon seeds could be reduced after 2 days of drainage, but the plants with 0.5ml and lmi alcohol treatment did not contain soluble protein. Flooded

The water has changed (Table 7), and the change in soluble protein content of plants that can be transferred to higher concentrations of alcohol-treated seeds is unlikely to be related to their tolerance to flooding. g 根种化# Papaya is secretive before and after flooding

The remaining X, W, V and the meanings represented are the same as those in Table 3.1 above. 28 1373304 The new white protein will accelerate the shouting of total free amino acids, and the ability of the egg to be inhibited or the anaerobic metabolism will cause the amine. The base acid stores a tendency to accumulate free amino acids. However, the total free amino acid change in this experiment 'the total free amino acid content of the leaves of papaya plants without seed treatment was significantly reduced 2 days after drainage, presumably because papaya itself is not resistant to flooding The root-and-leaf hybrid salt reductase activity is low, and the ability to synthesize amino acids is low, resulting in less significant or even less variation of papaya leaves and roots in submerged water. However, in terms of the concentration of the same treatment-treatment, the total free amino acid content of the leaves of the lmi alcohol-treated seeds was the highest when the water was flooded for 2 days, and after the drainage, 0.5W and 1 ml of the alcohol-treated seeds. The total free amino acid was also significantly higher than that of the (four) ml alcohol treatment and the control group. Therefore, from the results of this test, the soluble protein of the alcohol-treated plant was increased compared with the no-supplemented group (Table 7), indicating that the leaf free amine The increase in basal acid is related to protein synthesis. According to the experimental results of a comprehensive society, the # papaya seeds processed in addition to the seed (five bud rate) can also improve the flood-tolerance ability of the subsequent growing plants, and delay or reduce the damage caused by the flooding. For example, the changes of chlorophyll and chlorophyll in the leaves of the _ _ _ papaya plants are less in the flooding, indicating that the plants are flooded by the flood; the material, its Wei Weichi is high and the rate of acetaminophen is higher. Low, 2 by increasing the _dehydrogenase (ADH) 'to enable _ can _ sugary side to obtain the amount of monthly b and the plant resistance to k stress, peroxide peroxidation (10) activity, total phenolic compound content also Increased domain potential; flooded trees, transfer of carbohydrates, to increase the accumulation of Wang Ke * sex sugar, the powder of the temple is also an important adaptation of papaya plants to the adversity, the mineral nutrient element, nitrogen, helium, Potassium, _ has an increase trend 29 1373304 1 · 'potential' or change is not significant, trace elements such as iron, manganese, zinc, copper are reduced, can avoid toxic effects. The use of alcohol to treat papaya seeds, the seedlings of which are germinated are more tolerant to flooding, and can also be used to screen for flood-tolerant strains for reference in breeding new lines, and to enhance the growth of papaya for flooded environments. Adaptability, thereby enhancing or maintaining the yield and quality of papaya fruit. Other Detailed Descriptions All features disclosed in the specification can be combined in any combination. This statement can be replaced by the _, phase levy. Therefore, (4) Alternatives to the main-medical-purpose alternatives: The disclosed features are only examples of a series of J-like objects or similar features. Spears The person skilled in the art can determine the characteristics of the present invention and can be modified in the uncompressed SlT' to suit various uses and conditions as well as other specific aspects. The purpose of this application in the patent model [simplified description of the diagram] Figure 1 is the comparison of the seed surface, "it β

Photograph of the appearance change between the water and the quasi-water. Fig. 4 is a photograph showing the appearance change of the ‘(4), papaya after the seeds are treated with steam. $ Plants in flooding Figure 5 shows the situation of the seedling scale changes of the 'Tai Nong No. 2' before the flooding treatment. Figure 6 shows the change in the roots of the 'Tai Nong No. 2' papaya seeds treated with alcohol vapor before and after flooding. Figure 7 shows the change of the seedlings of the 'Tai Nong No. 2' papaya whose seeds were treated with alcohol vapor before and after flooding. Figure 8 shows the changes in alcohol dehydrogenase activity in the roots of 'Tai Nong No. 2 and Papaya plants' treated with alcohol vapor before and after flooding. [Main component symbol description] 31

Claims (1)

1. 1373304 VII. Patent application scope Supplementary correction replacement page is used for breeding (4) Method for jujube plants with flooding resistance, which is characterized by: (1) placing the JI fresh money nm at a concentration of 0.25 ml / 4L hair age / representing the volume of anhydrous alcohol added in the 4L beer suction), the anhydrous alcohol vapor volatilized + treated at 3CTC! Up to 3 days; (2) Seeding the alcohol-treated seeds to germinate; Papaya seedlings of the leaves are flooded for 2 days and then ^). Selecting a plant that grows well after drainage is a method for flooding papaya planting patents. The method used in step (1) = vapor inversion is α1! Wei's anhydrous alcohol 3. Post 1 item Tender, used in _) = refined Luo gas concentration of 05m fox to lmi / 4L of anhydrous alcohol, the method of claim (4), the method of wine processing in step (1) is 2 to 3 days. The method of applying for the patent remuneration item, in which the processing time for money is 3 days.广心酉楕32 4. 1373304 4 · « / Year of the Moon 6 曰 Supplementary Correction Replacement Page 6. - Viewing Gao Fa Lai County Reading Narrative _ Qian Li's method, which includes (4) Mu Lu Qiu Fa » Pre-requisite concentration is α25 secret礼至2 • The standard (representing the volume of anhydrous alcohol added to the 4L breathing tank) is volatilized into an anhydrous alcohol vapor for 1 to 3 days. 7. If you apply for the branch of item 6, the volatilization of the essence is 0.25 ml/4L to 1 ml/4L of anhydrous alcohol. #8·如(4) The method of item 6 of the patent face, which is volatilized by anhydrous alcohol with a vapor concentration of 0.5 mI/4L to 1 ml/4L. 9. The method of claim 6, wherein the steam is treated for 2 to 3 days. 1〇. f Please refer to the method of item 6 of the patent scope, wherein the treatment time of the alcohol vapor is 3 days. 11. As in the method of the cap patent paradigm, Wei-step comprises submerging the papaya seedling plants after sowing and germination of the alcohol-treated seeds. 12. If the method of claim 11 is applied, the flooding treatment is to flood the papaya seedlings for 2 days and then to collect water & days. ‘ 33