BE1029745B1 - Use of exogenous anhydrous D-Trehalose to improve the jointing and firmness characteristics of Oryza sativa l - Google Patents

Abstract

The present invention belongs to the technical field of oryza sativa l cultivation, and specifically discloses the use of exogenous anhydrous D-Thehalose to improve the filling and fruiting characteristics of large-type oryza sativa l ears. It was shown that anhydrous D-Trehalose treatment not only improved the photosynthetic assimilation capacity of oryza sativa l during the filling period, but also influenced the process of material accumulation and translocation and increased the accumulation of non-structural carbohydrates in the stem sheath during the tassel stage, thereby significantly altering the lower seed filling characteristics and increasing seed filling rate as well as seed quality.

Description

Use of exogenous anhydrous D-Trehalose to improve the jointing and firmness characteristics of Oryza sativa

Technical area

The present invention belongs to the field of Oryza sativa planting techniques | and describes in particular the use of an exogenous anhydrous D-Trehalose to improve the jointing and firmness characteristics of Oryza sativa 1 of the large ear type.

Core technology

Oryza sativa | (Oryza sativa L.) is one of the three most important food crops in the world and its demand continues to increase as the world population increases, so a greater amount of Oryza sativa 1 (Oryza sativa L.) can only be produced on limited land by increasing production per unit area. Increasing production of Oryza sativa | will effectively address the global food shortage, and the ups and downs of Oryza sativa 1 production depend on the volume size and filling degree of the slurry. Oryza sativa grain coulis | is an important stage in the growth and development of Oryza sativa 1 and a critical period for determining the yield of Oryza sativa |, affecting the firmness rate, grain weight, yield and quality of rice of Oryza sativa L. During this period, the products of photosynthesis are transported mainly in the form of sucrose from their "source" site of formation (leaves, stem sheath) to the "bank" (grains). But the big spike type Oryza sativa | variety tends to be strong, weak grain differentiation is obvious, in general, strong grains have the characteristics of rapid slurry, good filling and high grain weight; Weak grains, on the contrary, have the characteristics of slow slurry, poor filling and low grain weight, which has become a bottleneck for limiting production and improving quality rice. Therefore, to increase the production of Oryza sativa |, it is necessary to effectively increase the level of low particle slurry.

D-Trehalose anhydrous is derived from two glucose molecules a- Non-reducing disaccharides linked by 1,1 - glycosidic bonds, widely occurring in nature, are present in small amounts in plants. The metabolic pathway of

D-Trehalose anhydrous in plants is as follows: uridine diphosphate glucose and 6 - Phosphate Glucose first form D-Trehalose anhydrous - 6 - phosphate (t6p) under the action of D-Trehalose anhydrous - 6 - phosphate synthase, then are dephosphorylated by D-Trehalose anhydrous - 6 - phosphate phosphatase to form D-Trehalose anhydrous, which is finally hydrolyzed by the enzyme

D-Trehalose anhydrous to produce two glucose molecules.

D-Trehalose anhydrous acts as a signaling substance to regulate plant growth and development. As the endogenous anhydrous D-Trehalose content of the plant increases, the tóp content also increases. Tóp is widely distributed in plants and its signaling regulatory pathways may be directly involved in several plant physiological activities such as seed germination, seedling growth, flowering and aging. Studies have shown that tóp content in plants exists in proportion to sucrose content and is linked to starch metabolism.

However, current studies on the fastness of anhydrous D-Trehalose with grain slurry have not been reported.

Content of the invention

Considering the above technical problems, the invention proposes the following technical solutions:

The subject of the present invention is the use of exogenous anhydrous D - Trehalose to improve the grouting and stiffening characteristics of Oryza sativa | large ear type.

Preferably, said exogenous anhydrous D - Trehalose makes it possible to improve the optical contracting capabilities.

Preferably, said exogenous anhydrous D - Trehalose is capable of increasing the net rate of photosynthesis, STOMATIC conductivity and rate of transpiration, and reducing the intercellular concentration of carbon dioxide.

Preferably, said exogenous anhydrous D-Trehalose is capable of enhancing the accumulation and transport of non-structural carbohydrates in the stem sheath.

Preferably, said exogenous anhydrous D - Trehalose makes it possible to increase the rate of grouting of the grains and to shorten the active period of grouting.

Preferably, said exogenous anhydrous D - Trehalose makes it possible to improve the quality of the grains.

Preferably, the specific use of said exogenous anhydrous of D - Trehalose is as follows: an exogenous anhydrous of D - Trehalose at a concentration of 15 — 200 mmol | - 1 is sprayed foliarly in the middle and at the end of the gestation phase with a spray quantity of 200 ml.m - 1.

Preferably, said exogenous anhydrous D-Thehalose is sprayed at a concentration of 50 mmol-L-1.

The present invention also provides a spray for improving the filling and setting characteristics of oryza sativa | of the large ear type comprising

Exogenous anhydrous D-Trehalose as described in any of the above.

Compared to the prior art, the beneficial effects of the invention are:

The subject of the present invention is the use of exogenous anhydrous D - Trehalose to improve the grouting and stiffening characteristics of Oryza sativa | large ear type. Spray application of different concentrations of anhydrous D - Trehalose treatment not only improved the photocontractualization ability of Oryza sativa | at the grouting stage, but also affected the process of accumulation and transportation of the substance, enhancing the accumulation of non-structural carbohydrates in the rod sheath at the pumping stage, thereby clearly changing the grouting characteristics of the grains at low potential, increasing the grain jointing rate as well as the grain quality.

Description of accompanying drawings

Fig 1 is the evolution of the particle weight of the material to be tested for different concentrations of anhydrous D - Trehalose treatment; CO - control, clear water spray, C15, C30, C50, c70 and C100 respectively represent sprayed anhydrous D - Trehalose concentrations of 15 mmol | - 1.30 mmol 1 - 1.50 mmol | - 1.70 mmol | - 1 and 100 mmol.1 - 1; SS - strong grain; Is - weak particles;

Fig 2 is the evolution of the grouting speed of the test material for different concentrations of anhydrous D - Trehalose treatment; CO - control, clear water spray, C15, C30, C50, c70 and C100 respectively represent sprayed anhydrous D - Trehalose concentrations of 15 mmol | - 1.30 mmol | - 1.50 mmol | - 1.70 mmol | - 1 and 100 mmol.1 - 1; SS - strong grain; Is - weak grains.

Specific implementation arrangements

The invention is illustrated in more detail in relation to exemplary embodiments and the corresponding figures, the following exemplary embodiments being given for purely illustrative purposes and not limiting the scope of the invention.

The subject of the present invention is the use of exogenous anhydrous D - Trehalose to improve the grouting and stiffening characteristics of Oryza sativa | large ear type. More precisely, the invention studied the influence of exogenous anhydrosis

D - Trehalose on photosynthetic properties, accumulation and transport of stem sheath material, grain slurry characteristics and yield of Oryza sativa | large cob type by applying different concentrations of

D-Trehalose anhydrous foliar spray from material with significant differences in strong and weak grain slurry. The invention contributes to improving the solidity of the slurry of weakened grains of the large ear type Oryza sativa | and, therefore, the yield of the large ear type Oryza sativa L.

Example 1

Effect of exogenous anhydrous D-Trehalose on the yield of Oryza sativa | large spike type 1, test site and equipment

The trials were carried out in 2021 at the Izhong Test Base (31°28'55"N, 117°13"55"E) of Anhui Agricultural University, Guohe City, Lujiang County, Anhui Province. The test material is w1844 line of large ear rice with significant differences in strong and weak grain slurry. 5 2, test design

Large field planting, thin vase sowing method on plastic diskette for cultivating seedlings, sowing on May 23, 2021, transplanting on June 19, 2021. Row distance is 30.0 cm x 13.3 cm, 3 seedlings per cavity, cell size 10 m x 7 meters. The total fertilization is: urea 330 kg / hm2, superphosphate 450 kg / hm2, potassium chloride 300 kg / hm2. Urea is applied in the ratio of base fertilizer: tillering fertilizer: peak fertilizer = 3:3:4, superphosphate is used as base fertilizer and potassium chloride is applied in the ratio of base fertilizer: tillering fertilizer = 3:3:4. tip = 1:1. Other field managements are implemented according to the requirements of the local high yield crop.

A total of 6 treatments are implemented for this test: CO: spraying an equal amount of clean water as a control; C15: spray of 15 mmol: L - 1D-Trehalose anhydrous solution; C30: spray of 30 mmol: L - 1D-Trehalose anhydrous solution; C50: spray of 50 mmol: L - 1D-Trehalose anhydrous solution; C70: spray of 70 mmol: L-1D-Trehalose anhydrous solution; C100: — spray 100 mmol: L - 1D-Trehalose anhydrous solution. Foliar application was carried out in the middle and at the end of the gestation phase with a spray quantity of 200 ml: m - 2. 3, sampling method and determination indicators 3.1 sampling method

Oryza sativa | ear flowering period, from each treatment, select the ear registration marker of the main stem that flowers on the same day and the ear size is uniform, distinguishing strong and weak kernels , defining strong grains as glume flowers on the 3 primary branch peduncles in the upper part of the ear; Weak kernels are glume flowers on 3 primary pedicels at the base of the ear. 3.2 determination of yield and factors constituting yield

During the ripening period, 120 ears are randomly selected per cell, the rice ears are taken out and packed into mesh bags, the number of grains per ear, the weight of a thousand grains, the firmness rate and the performance are examined after air drying. The solidity rate and yield are calculated according to the following formula:

Firmness rate (%) = number of solid particles / total number of particles x 100%;

Yield (kg / mu) = number of ears per unit area (mu) x Number of grains per ear x Solidity rate (%) x Weight in thousands of grains (g) x 10 - 6. 5 Results

The results are presented in Table 1.

Table 1 Oryza sativa l yield and yield constituent factors for different concentrations of D-Trehalose anhydrous

Grain processing Production nt Grams Grains Grains Solid grains per (kg/Mu) strong-SS weak-IS strong-SS weak-IS pi [poor [maas maa | oan [eu enne 85.242.3° | 74.842.24 | 26.4+0.1° 23.340.19 | 244.33.84P | 703.9+26.644 gold maer maan | omen ne EEA 87.745.3be | 79.445.3° | 27.440.1P 23.9+0.2° | 267.747.512 c [meer (rate mar | re PTE" 88.6+1.7° | 82.241.7% | 28.2+0.1P 24.40.25 | 271.0+4.042; [er (ne moar [vaar nee] 92.940. | 83.6+0.7 " | 29.805 | 26.140.2* | 267.7+0.883 | 872.0+2.23° re [naar aa zoen amor er 89.5+1.9° | 80.641.8be | 281403) | 242401 | 260.043.052 | 784.9410.75° rare oee ara roer 87.242.5be | 79.842.4° | 27.840.2P 23.840.29d | 266.742.672 | 785.5+12.18°

Note: CO - control, spray with clean water; C15, C30, C50, c70 and C100 represent concentrations of 15 mmol respectively | - 1.30 mmol | - 1, 50 mmol.l - 1, 70 mmol.l - 1 and 100 mmol | - 1 of the pulverized anhydrosis solution.

Different lowercase letters after data in the same column indicate significant differences between treatments at the 0.05 level.

As shown in Table 1, the different concentrations of anhydrous treatment

D - Trehalose made it possible to significantly increase the yield of Oryza sativa, the increase in yield being the greatest with the C50 treatment, the lowest with the C30 and C15 treatments and the lowest with the c70 and C100 treatments. The efficiency of the C50 increased by 23.88% compared to CO. With different concentrations of D-Trehalose anhydrous, the stiffness ratio and thousand grain weight of strong and weak grains were improved, with the same change trend between different treatments as the yield. Among them, C50 processing has the highest rate of strong and weak grain toughness and thousand grain weight increase. Compared to CO, the strong and weak grains of

C50 showed an improvement of 9.04% and 11.76% in firmness and 12.88% and 12.02% in kilograin weight, respectively. The C50 treatment gives the best results.

Example 2

Effect of exogenous anhydrous D-Trehalose on grain slurry dynamics

The test equipment, test design, sampling method and data analysis method are the same as Example 1. 1. Sampling and determination method

The dynamics of grain slurry during the flowering period of Oryza cob — sativa | is selected from each treatment for the registration marks of the main stem spike that flowers on the same day and the size of the spike type is consistent. We distinguish between strong and weak grains, defining the strong grains as glume flowers on the 3 primary peduncles in the upper part of the ear and the weak grains as glume flowers on the 3 secondary peduncles at the base of the ear. ear. Sampling every 5 days after flowering until maturity. After killing the grain samples at 105 °C for 0.5 H, they were dried at 80 °C to constant weight for determination of grain slurry dynamics.

According to the method described in “Zhu Qingsen, Wang Zhichin, Zhang Xuan, et al. Oryza sativa | grain grouting Characteristics Study. Jiangsu Agricultural

College proceedings. 1995, 16 (2): 1 - 4.", the grain grouting process was adjusted using the Richards equation to calculate the characteristic parameters of the corresponding grouting (specific reference: Richards f. a flexible Growth Function for Empirical use. J Exp bot. 1959, 10 (29), 290 - 300). Richards equation:

HAS

W = —m —— (1 +Be)N

The Richards equation leads to the grout rate (R):

R AkBe*

Nl +B ek) IN+IJN

Where W is the particle weight (MG), a is the final particle weight (MG), t is the time after flowering (d) and B, K, N are the parameters determined by the regression equation. The main grout parameters are: starting grout potential Ro, maximum grout rate GRmax, average grout rate GRmeam, active grout period d, time to reach maximum grout rate Tmax. 2 the results

The Richards equations were fitted for grain reuse for each strong and weak grain slurry process, taking the number of days after flowering as the independent variable and grain mass as the dependent variable. The results are presented in Table 2 and Figure 1. The characteristic parameters of strong and weak grain grouts for different D-Trehalose anhydrous concentration treatments, calculated from the equation of

Richards, are presented in Table 3.

The mass gain of strong and weak grains varied significantly between treatments (Fig 1). The mass of strong grains is always greater than that of weak grains throughout the slurry, and grain mass increases rapidly after flowering, reaching a maximum around 35D after flowering, after which the variation in grain mass tends to decrease. stabilize, While weak grains slowly gain mass for a long period after flowering, reaching their maximum around 50 days after flowering. Spray treatment

D-Trehalose anhydrous changed the grain quality during slurrying, and the mass increase of vulnerable grains was improved compared to CO, whose mass increase curve is located above the d curve. increase in CO mass. Among these, the largest increase in grain mass was observed with the C50 treatment, with the mass increase curve being the highest.

Variation of grouting rate of strong and weak grains under different treatments as shown in fig 2, variety w1844 has a pronounced asynchronous grouting phenomenon, the grouting rate of strong grains quickly reaches the peak of grouting rate, and then decreases rapidly , while the grout rate of weaker grains increases slowly, the time to reach the peak of the grout rate is later, and the rate of stronger grout decreases more slowly after reaching the peak. The grout rates increased with each D-Trehalose anhydrous concentration treatment, where the maximum peak grout rate for C50-treated strong and weak grains was the highest compared to the control group.

Table 2 Characteristic parameters of grain slurry at different concentrations

D-Trehalose anhydrous treatment mee A IS PE TN TE

Note: CO - control, spray with clean water; C15, C30, C50, c70 and C100 represent concentrations of 15 mmol respectively | - 1.30 mmol | - 1.50 mmol.! - 1, 70 mmol.1 - 1 and 100 mmol.1 - 1 of the pulverized anhydrosis solution. SS - strong grain, Is - weak grain. TO - amount of terminal growth; B - Initial settings; K - Growth rate parameters; N - Shape parameters; R2 - decision factor.

As shown in Table 2, the results of the fitting equation between different treatments are R2 > 0.99, which indicates that the equation can objectively reflect the dynamics of grain slurry.

Table 3 Richards equation parameters for grain slurry process at different concentrations D-Trehalose anhydrous treatment pee ee Tao

Ro treatment (mg grain“d”) (d) | (mg’grain”) | (mg’grain“-d”) | (d)

Note: CO - control, spray with clean water; C15, C30, C50, c70 and C100 represent concentrations of 15 mmol respectively | - 1.30 mmol | - 1.50 mmol.! - 1, 70 mmol.1 - 1 and 100 mmol.1 - 1 of the pulverized anhydrosis solution. SS - strong grain; Is - weak grains. Ro - initial growth potential; GRmax - maximum grout rate; Tmax - time to reach the maximum grout rate; Wmax - particle weight at maximum grout rate; GRmean - average grout rate; D - active grout period.

It can be seen from Table 3 that the different anhydrous treatments of D - Trehalose at different concentrations have different effects on the different characteristic parameters. Compared with CO, the grout initiation potential (Ro), average grout rate (GRmean) versus maximum grout rate (GRmax) of strong and vulnerable grains treated by C50 were improved and the active grout period of vulnerable grains was shortened, that is, the actual grouting time was shortened, where the improvement of the grouting strength of vulnerable grains was more significant. With the C50 treatment, final growth was significantly improved compared to CO and other treatments. The active pouring period of each treatment is shorter than CO, but at the beginning of the grout has a higher growth potential, throughout the grout can maintain a higher pouring rate, so it can still achieve a higher particle mass.

Example 3

Effect of exogenous anhydrous D-Trehalose on photosynthesis in the sawing phase

The test equipment, test design, sampling method and data analysis method are the same as Example 1. 1, determination method

The net photosynthesis rate (PN), intercellular carbon dioxide concentration (Ci), STOMATICAL conductivity (Gs) and transpiration rate (Tr) of Oryza sativa leaves | were determined using a portable photosynthesizer

Li - 6400 from the American company Li - cor, during the sawing period of Oryza sativa |, selecting 10:00-11:00 am on a clear day, taking the average of 5 repeated trials. 2. results

The results are presented in Table 4.

Table 4 Photosynthetic characteristics at different concentrations D-Trehalose anhydrous treatment

Rate of Rate of

STOMATICAL TREATMENT | intercellular photosynthesis transpiration t Gs CO:

Pn (umol:m?:s%) Tr (mmol: ms) (mmol: m?s71}) Ci (umol mol!)

Note: photosynthetic data in the table are determined during the sawing period; CO - control, spraying with clean water. C15, C30, C50, C70 and

C100 respectively represent concentrations of 15 15 mmol-L*, 30 mmol-L**, 50 mmol Lt, 70 mmol Lt and 100 mmol:L* of the sprayed anhydrosis solution.

Different lowercase letters after data in the same column indicate significant differences between treatments at the 0.05 level.

Photosynthetic properties of Oryza sativa 1 leaves such as net photosynthesis rate (Pn), STOMATICAL conductivity (Gs), intercellular CO: concentration (Ci) and transpiration rates (Tr) were significantly changed at different concentrations of D-Trehalose anhydrous. As shown in Table 4, variable 616 vations of PN and GS were observed in leaves of Oryza sativa | treated with different concentrations of D-Trehalose anhydrous compared to the control. Among all treatments, the photosynthetic performance was highest with the C50 treatment, where Pn, Gs and Tr were significantly higher than the CO treatment, 33.49%, 60.00% and 9, respectively. .64%. Increasing the net rate of photosynthesis increases the accumulation of sugars, thus providing a more adequate “source” for the slurry.

Example 4

Effect of exogenous anhydrous D-Trehalose on pedicle material accumulation and transport

The test equipment, test design and data analysis method are the same as Example 1. 1. Method of sampling and determination

From flowering until 50 days after flowering, take 30 main stems 10 days apart until mature, divided into 3 repetitions. After 0.5 H of cyanicide at 105°C, dry at 80°C to constant weight and weigh, grind (100 mesh sieve), fractionate with self-labeled bags to determine the non-structural carbohydrate content of the stem sheath. The extraction, determination and calculation of non-structural carbohydrates from the stem sheath were carried out with reference to the method of “Yoshida S. Physical Aspects of Grain Yield. Annu Rev

Plant Physiol. 1972, 23, 437 - 464. », using the colorimetry method

Anthrone. And calculate the accumulation of non-structural carbohydrates (NSC) in the stem sheath, the transport of NSC in the stem sheath, the transport rate and the contribution to the grain at the hair removal stage compared to the maturation, according to the following formula:

Transport of NSC in the stem sheath = accumulation of NSC in the stem sheath at the spike stage - NSC residue in the stem sheath at the ripening stage

Stem sheath NSC transport rate (%) = stem sheath NSC transport / stem sheath NSC accumulation at spike stage x 100

Contribution (%) of stem sheath NSC to grain = stem sheath NSC transport / grain dry weight x 100 2 results

The results are presented in Table 5.

Table 5 accumulation and transport of substances in the sheaths of lower stems treated with D-Trehalose anhydrous at different concentrations

Road phase Period of NSC Volume | NSC rate

NSCTrate

NSC total maturation NSC of contribution

Traitem TM nsc at total transbordem | grain transshipment ent heading (g/m?) TM nsc at ent ent

C nsc maturity (g/m? ) TNsc TRnsc (%) (g/m°) (g/m?) 210.54+1.19 111.22+0.44 99.33+0.3f 47.18+0.59 13.42+0.4° 221.54+4. 1° 112.89+0.1° 108.65+0.69 | 49.04+0.3° 14.11+0.6° 232.82+0.52 109.61+0.4° 123.22+0.6P | 52.9240.4P 15.03+0.4e 236.18+0.3* 107.42+0.3f 128.7640.52 | 54.52+0.6* 15.51+0.74 227.06+0.2" 114.33+0.5" 112.46+0.7° | 49.53+0 4° 14.06+0.4°

C100 220.03+0.6° 115.39+0.42 104.64+0.7° | 47.56+0.59 13.59+0.24

Note: CO - control, spray with clean water. C15, C30, C50, c70 and C100 respectively represent concentrations of 15 mmol Lt, 30 mmol-L*, 50 mmol Lt, 70 mmol L and 100 mmol-L* of the sprayed anhydrosis solution.

Different lowercase letters after data in the same column indicate significant differences between treatments at the 0.05 level.

The accumulation and transport of stem sheath material under the treatment of different concentrations of D-Trehalose anhydrous are shown in Table 5.

As shown in the table, the different anhydrous treatments at different concentrations of D - Trehalose all enhanced the accumulation of NSCs in the stem sheaths at the spike stage, and the accumulation of NSCs in the stem sheaths showed a tendency to C50 > C30 > c70 > C15 > C100 > CO. The highest NSC accumulation in the stem sheath during the sawing phase was observed under the C50 treatment, with a 12.18% increase in NSC accumulation in the stem sheath. the stem compared to the control, while treatment with different concentrations of D-Trehalose anhydrous resulted in a significant increase in the transport of the substance from the stem sheath compared to the control. Among these, C50 treatment has the highest transshipment rate than other treatments. Compared to the control, the grain contribution was significantly higher for each concentration treatment than for the control, while that of the C50 treatment was still significantly higher than that of the control. other treatments.

The examples above are given only by way of illustration and not limitation of technical solutions of the invention; Although the invention has been detailed with reference to a more advantageous embodiment, those skilled in the art in his field should understand that it is still possible to modify particular embodiments of the invention or to substitute in a manner equivalent certain technical characteristics; Without departing from the spirit of the technical solution of the invention, this must be covered by the scope of the technical solution for which protection of the invention is requested.

Claims (9)

Claims 1. Use of the exogenous anhydrous D-Trehalose in improving the jointing and robustness characteristics of Oryza sativa | large ear type. 2. Use according to claim 1, characterized in that the exogenous anhydrous D-Trehalose makes it possible to improve the optical contracting capacity. 3. Use according to claim 2, characterized in that the exogenous anhydrous D-Trehalose is capable of increasing the net rate of photosynthesis, STOMATIC conductivity and transpiration rates and reducing the intercellular concentration of carbon dioxide. 4 Use according to claim 1, characterized in that the exogenous anhydrous D-Trehalose is capable of improving the accumulation and transport of non-structural carbohydrates in the stem sheath. 5. Use according to claim 1, characterized in that the exogenous anhydrous D-Trehalose makes it possible to increase the rate of grouting of the grains and to shorten the period of active grouting. 6. Use according to claim 1, characterized in that the exogenous anhydrous D-Trehalose makes it possible to improve the quality of the grains. 7. Use according to claim 1, characterized in that the specific method of using the exogenous anhydrous D-Trehalose is as follows: foliar spraying of the exogenous anhydrous D-Trehalose at a concentration of 15 to 200 mmol Lt at middle and end of the gestation phase with a spray quantity of 200 mlm". 8. Use according to claim 7, characterized in that the exogenous D-Trehalose anhydrose has a spray concentration of 50 50 mmol-L"!. 9. Spray agent for improving the jointing and robustness characteristics of Oryza sativa | of large ear type, characterized in that it comprises exogenous anhydrous D-Trehalose according to any one of claims 1 to 8.