JPS59179017A - Application of specific illumination treatment for increasing antocyanine in economically important crop - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention involves the use of specific lighting treatments to enhance anthocyanin formation in economically important fruit and ornamental crops without affecting other quality characteristics and growth. It is therefore a general object of the present invention to provide such a new and improved method.

Anthocyanins are water-soluble pigments that are responsible for the attractive colors of flowers, leaves, and fruits. Apart from its biological role, the formation and stability of the andocyanins are of economic and regulatory importance as they have implications for the marketability of crop products.

In field and greenhouse agriculture, red color enhancement of crops has been achieved by spraying the entire crop or specific parts of the crop with a chemical conditioning agent. Also,
In some cases, genetic selection and breeding methods have been used to improve coloration.

However, the chemical moderation agents traditionally used by growers to ensure a timely red coloration in a variety of ornamental and fruit crops have had undesirable side effects such as leaf drop, shortened shelf life, and root damage. (e.g., suppression) and often exhibited significant variability in response. Genetic selection and breeding strategies are laborious and time consuming.

It is known that anthocyanin synthesis in various tissues and plant species is promoted by light.

This photostimulation involves 1) a low-energy red/far-infrared reversible phytochrome-regulated reaction and 2) a high-irradiation reaction (HIR) that is most effective in the blue and far-infrared regions of the visible light spectrum.
This appears to be achieved through at least two photochemical reactions. II 1:R of andocyanin accumulation is usually f'J\ and has been studied as a phytochrome or other unknown photoreceptor.

One aim of the present invention is to promote anthocyanin formation in fruits or ornamental crops without causing any harmful conditions.

The fruits of the seeds and the kernels, from which the ornamental crops turn red in time, have great economic significance in terms of their production and marketing. There are many factors that influence anthocyanin formation, one of which is the original influence. The effects of light on fruits and ornamental crops were studied using a variety of strategies. for example,
Ita-green mature apples and/or lingonberries in customary refrigerated conditions were exposed to blue light (10.82 mW/cm') and red light (10.82 mW/cm') and red light (with center points of maximum emission peaks at approximately 448 nm and 660 nm K, respectively) at various time intervals. 0.50m
The results showed a significant (over 46% on average over blue or red light treatment alone) enhancement in anthocyanin formation. Similarly,
Post-harvest illumination with red and blue light at 10°C also resulted in a significant (on average over 65%) enhancement in anthocyanin formation in mature apple skins compared to the vermilion-illuminated control group.

According to the present invention, anthocyanin formation in fruits or other crops can be promoted by subjecting them to a combined blue and red light treatment. crops,
IR up to 40 days prior to harvest, intensity range 1-2
00μw/crt? high intensity discharge and/or VH
O can be exposed to narrow band fluorescent lamps for 1-4 hr per day. It is preferable to expose apples to both blue and red light before harvest, but poinsettias should only be exposed to red light. After harvest, apples kept in a refrigerated state can be exposed to red light or red and blue light sequentially for 4 days.

Other objects, benefits and 'ffv!' of the invention! , υ, will become clear from the following description.

EXAMPLE Field experiments on various crops and studies using in vitro systems have shown that red coloration can be promoted through effective light exposure.

(1) Overview: (a) relative efficacy of various irradiances and spectral regions; (b) red/far-infrared convertibility and interrelationships; and (C) phytochrome involvement and photosynthesis on red medium HIR response. In order to investigate the possible involvement of anthocyanin formation, in vitro threads of selected model crops such as bilberry, apple and poinsettia were used to study anthocyanin formation.

(2) Grain materials and methods As crop materials, Vaccinium maerucarl, Vaccinium maerOcarl
On) AIT (obtained from Ocean Spray Company, Middleboro, Mass.): Poinsettia, Niphorbia pulcherrima (Eup
hor-biapulcherrima) J V-
14 (obtained from Nikke Nurseries, Incinatas, California); Oyohiringo 'Maquintas', MaIus domestica
ca ) (obtained from the University of Massachusetts Hoata Cultural Research Center, Belchertown, KY and Standard Oachaz, Hudson, MA)
was used. In most cases, apple skin tissue and poinsettia leaf tissue used for experiments were cut into uniform discs (α5 cm diameter) using a spring-loaded plunger. The discs cut and obtained from each organizational group were
The mixture was poured directly into methanol, immediately frozen in liquid nitrogen, and stored in a freezer.

Experiments were conducted in an environmental chamber (Condrold Invironments Ltd., Winnabeg, Canada) divided into five light-tight modules. A light fixture was placed across the top of each module, and the light intensity was modulated by adjusting the distance between the light fixture and the tissue. Keep each tissue at 25°C-27°C and separately store 371-7
40 nm l'n19: Narrow band light of wavelength (irradiance range - 0.01 mW/ls2 to 2 mW/crn
2) in a continuous manner each day.

Wavelength 371, 420, 448, 467, 504, 550
, 590, 660, and 74 DHm (supplied by GTB Sylvania Lighting Products, Denver, Mass.) with a 5 mil thick weather-resistant polyester film (supplied by GTB Sylvania Lighting Products, Denvers, Mass.). Martin Processing Company) (excluding 571 nm lamps). In addition, to absorb visible mercury lines (not in the adjacent spectral region of the narrowband emitter), the Uv
A plastic filter surrounding the prefilter was used. The bandwidth and filters used for each light source are known in the art. Methanol-HCI (99:1
Anthocyanins were extracted from whole tissues using the following methods. The extract was clarified by p-filtration and dilution of the extract with metal-HCI was performed within each set of plant tissues until the absorbance of the solution was read spectrophotometrically at 530 nm and 657 nm. In order to exclude the contribution of chlorophyll and its degraded products in the acid solution to the absorbance at 530 nm, the formula (A330'''0
33A657) was used.

(3) Results The action spectra on anthocyanin formation were measured for the fruit skins of bilberry and apple, and the discs of improved poinsettia obtained by Hamyo et al. The measurement of this action spectrum was carried out during the linear formation phase of anthocyanins. Representative results obtained from several action spectrum experiments are shown in Figure 1. This figure illustrates crop spectra for anthocyanin formation of selected model crops.

The disc is exposed to light of different wavelengths. 1M sucrose K
[L, Incubated. Anthocyanin formation was plotted as a function of the wavelength of light used in each incubation. Each point on this plot represents the average value of 15 samples. Each sample consisted of a 5° disc.

Cranberry fruit skin discs cultured in 0.1 M sucrose under light with wavelengths of 0.5 to 6.5 nm show two peaks of anthocyanin biosynthesis, a lower peak at 448 nm and a lower peak at 6.
It showed a higher peak at 60 nm. The spectrum of action of poinsettia leaf discs and apple fruit skin discs for anthocyanin formation was essentially the same as that of bilberry. However, the most effective light wavelength for anthocyanin formation in apple discs is
The backlight had a maximum emission peak of 448 nmVc.

The spectral stability for anthocyanin formation in selected plant species exposed to continuous blue or red light depends on the irradiance and length of the light. In the fruit skin discs of climbing bilberries and apples, anthocyanin synthesis occurs at 0% under backlight.
．． 82 m W/cm” and 1.19 m under red light
It is fully saturated with irradiation of ``W/c+++'' (Figure 2).
Figure 3 shows the effect of light intensity on anthocyanin formation in fruit skin discs. Discs cut from the fruit skin were immediately transferred into an incubation medium and exposed to different intensities of red light (1) and blue light (2) for 144 hours.

The value for each point is the statistical error of five separate experiments performed in triplicate.

For poinsettia leaf discs, the blue light intensity required for saturation of anthocyanin formation was the same as that for apple skin discs. However, the saturation of red light intensity for anthocyanin formation was approximately one-fourth of that required in apple fruit skin discs (Figure 3). Figure 6 shows
Figure 3 shows the effect of light intensity on anthocyanin formation in poinsettia leaf discs. Discs cut from improved leaves were immediately transferred to incubation medium and exposed to different intensities of red and blue light separately for 120 hr.

The value at each point is the average mean statistical error of the values of five separate experiments performed in triplicate.

Regarding the time course of anthocyanin synthesis under saturated blue light and red light in poinsettia leaf discs.
Facts about anthocyanin synthesis: Approximately 12 hours without any visible results
It was found that there is an initial lag phase of r. The onset of anthocyanin formation is after this lag stage, and anthocyanin formation reaches steady state by 120 hr and 216 hr of red and blue light treatment (Figure 4).

The fourth FA shows the time course involved in anthocyanin synthesis in Poinsettia leaf discs. Immediately transfer discs cut from leaves of improved Boinsetea to incubation medium and incubate with red light (660 nm, 0.30 nl).
W/CIn and blue light (448nm, 0.82mW
/□2), and then harvested at the times shown in the table and evaluated for anthocyanin content.

The value at each point is the statistical standard error of the mean of five separate experimental values performed in triplicate.

As is clear from Figure 5, which shows the time course of the synthesis, anthocyanin synthesis in apple fruit skin discs is as follows:
Under saturated blue and red light, they commonly exhibited an initial lag of about 24 hours and reached steady state by about 144 hr and 196 hr, respectively. Discs cut from the fruit skin were immediately transferred to incubation medium and exposed to blue light (448n).
+n, 0.82mW/c1n”) and red light (660n
m, 119 mW/cnr9 at Ohr, harvested at the times indicated in the table and subjected to J-12 determination of anthocyanin content.

The value at each point is the statistical standard error of the mean of five separate experimental values performed in triplicate.

Since the action spectra for anthocyanin formation in apple and poinsettia showed maxima at light wavelengths of approximately 448 nm and 660 nm, the relative roles of these wavelengths in anthocyanin synthesis were tested. The data in Table 1 show the effect of blue light and red light interaction on anthocyanin synthesis in apples. Continuous if saturating light intensity
Apple fruit skin discs exposed to light formed more andocyanin than when exposed to continuous red light. However, when exposed simultaneously to continuous blue light and continuous red light, approximately 36% more anthocyanins were formed than continuous blue light alone. A similar effect was observed for continuous blue light when low red light flux was co-presented throughout the irradiation period. These results show that red light serves as a trigger for blue light, which functions through a "high irradiance response." However, the reverse is not true either. This is because pulses of blue light layered over continuous red light have not been equally effective.

Table 1 The interaction of the two light wavelengths on anthocyanin formation in poinsettia leaf discs was somewhat different.

As shown in Table 2, the most effective obscene band was the red light peak at 660 nm. 6 in anthocyanin formation in confectionery discs when continuous blue light was supplied together with continuous red light.
An increase of more than 0% was observed.

When short M cod was exposed to blue light and continuous red light, anthocyanin formation was equal to that of continuous blue and red light.

Table 2 The difference in the effects of red and blue light on anthocyanin formation appears to be related to the stability of photoreceptors (possibly phytochromes). In apple discs, photoreceptors appear to be relatively unstable, as brief exposures to red light are required between periods of exposure to blue light. Red light alone activates the phytochromes necessary for the blue high-irradiance response (HIR), making HIR possible, albeit at a lower level of efficiency. This effect probably provides some precursors necessary for andocyanin formation. Blue light reduces the levels of a unique inhibitor called phenylalanine ammonia-lyase, which interferes with the key enzyme involved in anthocyanin biosynthesis. It seems to be.

However, established criteria should be met before a phytochrome can be claimed to be relevant to a plant system. Table 3 Achievement of criteria regarding andocyanin formation
Shown below.

Table 3 Anthocyanin formation is short (10, nin) each day
The effect was completely abolished by immediate and subsequent exposure to 10 m1 of far-infrared (FR) light. Induction by low irradiance treatment and red/infrared cell inversion reactions provided evidence that at least one of the photoreceptors involved in anthocyanin formation is a phytochrome.

As shown in Table 4, the reciprocal changes in irradiance and irradiance j demonstrate that anthocyanin formation in poinsettia discs follows a reciprocal relationship, and that this response is a function of radiation rather than a function of irradiance alone. fil (I
X t ): Proved. This robust correlation indicates that only one photoreceptor is involved in the photoregulation of anthocyanin synthesis.

Table 4. Relatively few anthocyanin signatures form in response to short-term irradiation; long-term exposure to red light is required to obtain maximum formation evidence (Table 5). n11 responses were low energy;
Identified as a red/far-infrared reversible phytochrome reaction. Since the latter is recognized as a high-energy reaction, it is also called the wound irradiance response system of plant photoforms. The latter response suggested the dependence of phytochrome interaction on exposure time, or the existence of a photosystem (particularly photosynthesis) separate from phytochrome.

Table 5 To determine whether photosynthesis contributes to red light-mediated HIR responses by enhancing anthocyanin formation in poinsettia discs, studies were performed using various inhibitors of photosynthetic phosphorylation and chlorophyll synthesis. Table 6 shows the impact of cyclic and acyclic photosynthetic inhibitors on anthocyanin synthesis. poinsettia leaf disc,
Incubate with four separate inhibitors under 660 nm light for a period of time. 3-(3,4-dichlorophenyl)-1-dimethylurea (DCMU), acyclic photosynthetic phosphorylation of ammonium sulfate (NH4)2S04
, dinitrophenol (DNP), and antimycin-A (ANT-A), none of the inhibitors inhibited photo-mediated anthocyanin formation.

Cloth 6 Similarly, streptomycin (STM) and chloramphae, which are inhibitors of chloroplast appearance and chlorophyll synthesis,
Cole (CHP) is 6 degrees 10 ppm and 10 o
ppm, there is no shadow on andocyanin synthesis (Table 7).

Table 7. Basic characteristics of phytochrome responses such as relative potency of each ladder irradiance, red/far infrared reversibility, and reliable mutual flash association are unaffected by antibiotics.

The ratio of the amount of anthocyanins formed after 10 m1n of red light exposure and the amount of anthocyanins formed after 10 m1n of red/10 min of far-infrared treatment was the same in the inhibitor-containing incubation medium as that observed in the control. There was (Table 8). This discovery suggests that photosynthesis plays a role in red light-dependent anthocyanin formation.What is the effect of red light on synthesis and on the appearance of photosynthesis? They are unrelated to each other.

Table 8 (4) Discussion Vine: 17 The action spectra of anthocyanin formation in woolly peas, apple fruit skin, and poinsettia leaf discs showed one peak in the blue region of the visible spectrum and one peak in Akagi (Figure 1). . The irradiance dependence and spectral sensitivity of anthocyanin synthesis in tissues exposed to continuous irradiation depends on the exposure length (Figures 4 and 5). Thus, anthocyanin synthesis is controlled by a high irradiance response driven through phytochrome and other HIR photoreceptor interactions.

Red light was effective in stimulating anthocyanin formation, but this effect was abolished when red light was immediately followed by far-infrared light exposure. Strong reversibility was obtained with short exposures RtMj. This clearly indicates that phytochrome is involved (Table 3
).

Field research (1) Apple a, apples on trees treated with night shade and loss of light (varieties ``Maquintash'', ``Red
Delicious"), NQiJ40 days iM4 night blackout treatment (high intensity discharge and/or V HO narrow band fluorescent lamp; 1 pW/crl ~ 2001 tW/crl,
When exposed to the control group (no continuous exposure to light at night), an increase in anthocyanin formation was observed in one bite.

Apple varieties Makintash (1977 and 1978) and Red in the Washington and California rivers.
Comparative data for exposed vs. exposed material for Delicious (1978-1980) is listed in Table IK below. This table is
This figure shows the effect of night-time darkness treatment on the red coloration of apples at the time of harvest. It should be noted that in 1977, the trees were harvested 1) iJ and exposed to continuous light for 30 days.
The red coloring rate of apples increased from 738% to 78.3%4
．． This shows an increase of 5%. Also, in 1978, the group exposed to 30 days lost 62.8%, an increase of 98% compared to the 56% control.

There are numerical differences between years due to temperature, humidity, rainfall edge,
This is partly due to the fact that both seasons are not the same in terms of the degree of infiltration of numbers.

The applied light was a combination of continuous blue light and continuous early spring light with maximum emission peaks at approximately 448 nm and 66 Orlm, respectively.

Apple cultivars Red Delicious were grown in 1978 from Tsuenatchee, Washington and from Sunden, California in 1979, and were grown at 15 m1n over a 45 day period.
When exposed to heat light, the coloring of the /carriages was improved by 92% and 7%, respectively, compared to the control. When this trial run was repeated the following year and exposed for 40 days, the results were 92% and 12%, respectively.
showed an improvement of 5%.

1979 apple from Tsuenatchee, Washington (variety:
The effects of night shade interruption treatment on the growth and quality of "Red Delicious") at the time of harvest are listed in the following table ■. This table shows a comparison of apples treated with nocturnal or axillary treatment at two radiation doses and a control.

In particular, Table 3 shows that the night-shade interrupted lighting group had higher red coloration rate, as well as the total solid content (%) and dry weight of apples, than both the control and armpit treatment groups.

In contrast, the chemical growth Q-clearing ester, which is used commercially for the red coloration of apples, produces apples with poor storage properties, whereas apples subjected to Gyin-disruption treatment with blue and red light produce apples with poor storage properties. showed good storage quality characteristics. This is listed in the following table ■.

Table ■ Effect of night-shade interruption irradiation treatment on the quality of 1980 harvested apple fruit (variety [Red Delicious] - Top Red) (produced in Sunden, California) The treatment is a "control" that does not undergo this treatment.
It had various benefits compared to apples. As shown in the following Table IV, the irradiated apples were compared to the control object, solid content ($).
, weight, height, and red coloration rate, and also showed an increase in trunk length, making it more desirable for consumers (U, S. Extra Fancy or Fancy).

Apples treated with continuous light exposure retained the firmness of the "control" apples, had the good storage properties of the control apples, had a higher solids content, and had a higher percentage of red coloration than the control apples. In contrast, apples treated with ethrel had lower firmness, lower solids content, and poor storage properties.

This is shown in the following table ■.

b. Table V1 shows the red coloration rate of the 1979 Sunden, California apple "Red Delicious" under continuous post-harvest exposure control conditions and exposure conditions. Prior to harvesting, the exposed apples were exposed to approximately 100 μW/d of continuous light (emission peak: 66 onm) and blue (emission peak: 448 nm).
Exposure time (10:00 PM to 2:00 AM). After harvesting, the exposed apples were exposed to 660 nm light (approximately 1 o pw/c
rl) for 4 consecutive days, during which time the apples were kept refrigerated.

Table ■ 1979 "Red Delicious" apple grown in California, California.
0 days, continuous dark exposure treatment (HID and narrow band fluorescent lamp: 1 μW/c11-200) tw7cr+,
1 to 4 br/day), an increase in anthocyanin formation was observed compared to the control group (no continuous exposure to light at night) (Table ■).

Fruit growth (size) and quality (pulp firmness, solids content and shelf life) at harvest were not adversely affected by the night-interrupted light exposure treatment. Similarly, nuchal germination and fruit bud development were normal.

Table ■ Conclusion The use of specific light exposure systems (either in storage conditions or in greenhouse or field conditions) can considerably aid in color improvement of fruit and ornamental crops, while also inhibiting the growth or development of the tree from any harmful substances, including phytotoxins. It should not have any negative impact.

Thus, throughout the practice of this invention, under customary refrigeration conditions, greenhouse and field conditions, and using specific light exposure treatments, fruit coloration did not indicate any adverse effect on plant growth or development. It could have been improved without any problems. That is, the original storage variety characteristics of the fruit could be maintained, and there was no environmental pollution caused by dangerous chemical residues. Apples removed from conventional refrigeration were subjected to light exposure treatments at both pre-harvest and post-harvest times without subsequent fading.

[Brief explanation of the drawing]

FIG. 1 is an action spectrum of anthocyanin formation in selected model crops when discs of the crop are incubated in 0.1 M sucrose under various wavelengths of light. No. 21: J represents climbing berry (C) and apple fruit (
4) Chart illustrating the effect of light intensity on anthocyanin formation in the disc. FIG. 6 is a chart illustrating the effect of light intensity on anthocyanin formation in poinsettia leaf discs. FIG. 4 is a time course of anthocyanin synthesis in poinsettia leaf discs. FIG. 5 is a time course of anthocyanin synthesis in apple fruit discs. Procedural amendment: C. April 27, 1980 [I. Commissioner of the Patent Office Kazuo Wakasugi Display of the case Patent Application No. 49588 of 1988 is related to the person making the amendment -111 Patent applicant's representative Telephone number 273-6436 No. 41 Address 1. Subject of amendment: Inventive power of the specification, which is subject to amendment. I will correct it as written in Shochuhiro. 1. The scope of claims is amended as follows. ``Claims: (1) A method for enhancing anthocyanin formation in a product selected from the group consisting of fruits and crops, comprising subjecting said product to a combined blue light and red light treatment. , apple, grape, climbing berry and poinsettia. (3) The product group comprises apple varieties "Red Delicious", "Maquintas" and Malus domesticus. Power(
'Vaceinium macrocarp', grape variety 'Enhera', 'Vaceinium macrocarp'
on) AI T, as well as the poinsettia Euphorbia pulc
herrima) V-14. (4) The method described in item 1 of the scope of patent i4, wherein the product is subjected to a night shade treatment for several days prior to harvesting. 4. The method according to claim 4. (6) Perform night blackout treatment using a luminous material selected from the group consisting of luminance discharge and fluorescence at 1 to 200 μW/
cm” intensity for 1 to 4 hr per day,
The method according to claim 4. (A product selected from the group consisting of Apple Red Delicious, Macintash, Cranberry, Emperor Grape, and Poinsettia) under blue and red light with center points of harvest at 448 nm and 660 nm, respectively. A method for enhancing the formation of anthocyanins in the product, the method comprising: selecting from the group consisting of: (8) the product being poinsettia; The method according to claim 7, wherein the red light has the most luminous peak at Lighting is approximately 660 Bm
Red light with the center point of the maximum luminescence peak at about 448
If from the group consisting of a combination of blue light and red light having the center points of maximum emission peaks at 660 nm and 660 nm, respectively, if
8. The method of claim 7, wherein f is disclosed. 00) While keeping the harvested apples in the usual refrigerated state,
A method of enhancing anthocyanin formation in apples comprising continuous exposure to red light having a maximum emission peak centered at about 660 nm. 11. The method according to claim 10, wherein apples are exposed for four consecutive days. 2. In the 6th line from the bottom of page 21 (the last line in the footnotes of "Table 4"), the phrase "is." has been corrected to "±statistics standard deviation.". In the 6th line from the bottom of page 39, the phrase "after harvest" has been corrected to "after harvest, which continues for 50 days following the treatment shown in the following table ■."

Claims (11)

[Claims] (1) A method for enhancing anthocyanin formation in a product selected from the group consisting of fruits and crops, comprising subjecting said product to a combined blue light and red light treatment. 2. The method of claim 1, wherein the product is selected from the group consisting of apples, grapes, bilberries, and poinsettias. (3) The product group is the apple variety “Red Delicious”
, ``Maquintash'' and Malus domestica (MB
IuS domestica), grape variety "Enhera"), vineberry vaccinium macro+4rpon
) AIT, as well as Boinsetea's knee 7 Olbia
Euphorbia pujcherri
ma) The method of claim 2, comprising V-14. (4) The method according to claim 1, wherein the product is subjected to a treatment for several days prior to harvesting. (5) The method according to claim 4, wherein the product is subjected to a 40-day night shade treatment prior to harvesting. (6) The nighttime blackout treatment is carried out using a luminous material selected from the group consisting of high-intensity discharge and fluorescence at an intensity of 1 to 200 μWΔ for 1 to 4 hours per day. Method described. (7) Before harvest, products selected from the group consisting of Red Delicious apples, Macintosh apples, Emperor berries, and Poinsettia grapes were fertilized at various time intervals to find the center point of the maximum luminescence peak, respectively. 44
A method for enhancing anthocyanin formation in said product, comprising subjecting it to night-time interrupted illumination with a combined treatment of blue and red light at 8 nm and 660 nm. (8) The method of claim 7, wherein the product is poinsettia and the night-time interruption illumination is red light with a maximum emission peak at about 660 nm. (9) The M object is approached by a swarm of apples and grapes, and the night-shade interruption illumination produces red light with the center point of the maximum emission peak at about 660 nm, and about 448 m and 66 m.
8. The method of claim 7, wherein the light is selected from the group consisting of a combination of blue light and red light, each having a center point of maximum emission peak at 0 nm. (10) A method for enhancing the formation of anthocyanins in apples, which comprises continuously exposing harvested apples to red light having a maximum luminescence peak centered at about 66 onm while keeping the apples under normal refrigeration. (11) The method described in Item 10 of Patent Application No. 11 No. 7, wherein the phosphor is exposed continuously for 4 days.