Ecology

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Site descriptionThe experimental was conducted in Gengzhuang Town, Haicheng(40° 48′ N, 122° 37′ E), Liaoning Province from 2019 to 2020. This area is belonged to the continental monsoon climate zone of warm temperate zone, the annual average temperature is above 10 °C, the annual accumulated temperature is 3000–3100 °C, the frost-free period is about 170 days, and the annual rainfall is 600–800 mm. The soil at the experimental site is classified as brown earth29. Before the experiment, this test field had been in rotary tillage mode each year, with no straw return. The concentration of soil organic carbon, total nitrogen, available nitrogen, available phosphorus, available potassium, and soil bulk density in 0–20 cm surface layer were 12.50 g kg−1, 0.89 g kg−1, 129.6 mg kg−1, 25.96 mg kg−1, 117.94 mg kg−1, and 1.53 g cm−3, respectively. The components and nutrient contents of corn straw were shown in Table 1, The average temperature and precipitation from May 2019 to May 2020 are shown in Fig. 1.Table 1 Initial component content of corn straw.Full size tableFigure 1Daily precipitation and mean air temperature during the straw decomposition period from May 2019 to May 2020.Full size imageExperimental design and managementWe adopted a split plot design, with the main plot as the cultivation method, and with three cultivation options: no-tillage, deep loosening + deep rotary tillage and rotary tillage. Then, when adding straw as a decomposer, two methods were used: adding straw decomposer and not adding it. Our experimental approach included six treatments: No-tillage and straw mulching to the field + straw decomposer (NT + S); no-tillage and straw mulching to the field + no straw decomposer (NT); rotary tillage and straw mixed into the soil + straw decomposer (RT + S); rotary tillage and straw mixed into the soil + no straw decomposer (RT); deep loosening + deep rotary tillage and straw return to the field + straw decomposer (PT + S); and deep loosening + deep rotary tillage and straw return to the field + no straw decomposer (PT). Each treatment was replicated 3 times, located in random blocks, with a total plot area of 68.4 m2. At the same time as the previous year’s corn harvest, straw was returned to the field, crushed to about 10 cm long, spread evenly on the ground. No-tillage mulching and straw return to the field is the direct no-tillage maize sowing operation in spring; In the deep loosening + deep rotary tillage treatment, the soil is turned using a subsoiler to a depth of 35 cm, and then the straw is mixed into the soil through deep rotary tillage (a depth of 30 cm); and rotary tillage involves mixing straw into the soil with a rotary tiller to a depth of 20 cm and then raking it flat.Using the nylon net bag method (mesh bags were 5 cm × 6 cm, small; 15 cm × 20 cm, medium; and large, 25 cm × 35 cm; each size with an aperture of 100 mesh), we simulated three return modes. Soil added to the net bags was taken from the top 0–20 cm prior to sowing in 2019, in the corresponding plots of each treatment. Corn stalks were added at a ratio of 5:4 per stem and leaf (dry weight of stem and leaf of corn stalks in mature stage), and crushed to 2 cm long. In no-tillage treatment, 10 g straw was added to the medium mesh bag, in the rotary tillage and deep loosening + deep rotary tillage treatment, 10 g straw was evenly divided into five parts and put into five small net bags, then the five small net bags were evenly mixed into the soil of the outer large net bag and sealed, the weight of soil added to each large net bag is 2 kg, and the compactness between the inner net bag and the soil in the outer net bag was adjusted.Net bag layout was determined according to different treatment tillage patterns, and bags were placed in the field on seeding day in 2019. Deep loosening + deep rotary tillage was achieved by ploughing furrows 30 cm long, 15 cm wide, and 35 cm deep between corn rows in corresponding plots, large net bags were buried vertically in the furrows, filled with soil and compacted, so that return depth and straw distribution were basically the same as deep loosening + deep rotary tillage in the field. Rotary tillage mode was achieved by ploughing furrows 30 cm long, 15 cm wide, and 20 cm deep between corn rows in the corresponding treatment plot, the packed net bags were tilted in the furrows, filled with soil and moderately compacted, the top end of the net bags was level with the ground surface, which is basically consistent with the return depth of rotary tillage and straw distribution in actual field production. No-tillage mulching treatments involved laying the net bags containing straw on the ground and covering the four corners with soil to prevent the net bag from being blown away by the wind. The decomposer addition treatments involved evenly spraying c. 6.5 ml straw decomposer on the straw surface before bagging, in the treatment without decomposer, 6.5 ml water was sprayed on the surface of straw to maintain the same water content.In all treatments we applied the same amount of N, P and K (N 240 kg hm−2, P2O5 74 kg hm−2 and K2O 89 kg hm−2). The nitrogen fertilizer was urea, the phosphate fertilizer superphosphate, and the potassium fertilizer, potassium chloride. The brand of straw decomposing agent is Gainby and the model number is d-68 (created by NORDOX company and produced by Beijing Shifang Biotechnology Co., Ltd.). Straw decomposer dosage was 1.5 kg hm−2, diluted with water 100 times, and the effective viable bacteria number was ≥ 50 million g−1. The effective bacteria in the decomposer include: Bacillus licheniformis, Aspergillus niger and Saccharomyces cerevisiae and so on.Sampling and analysis methodsOn the 15th, 35th, 55th, 75th, 95th, 145th and 365th day after the nylon net bags were placed in the field plots, 3 bags were randomly sampled from each plot. For each net bag, we first washed the surface soil off with tap water, then washed the sample with distilled water 3 times, dried it at 60 °C, weighed it and then ground it to deter-mine the decomposition rate of straw and its components. At the same time, in the no-tillage treatments, 200 g soil was taken from 0 to 5 cm below the straw net bag, in rotary tillage and deep loosening + deep rotary tillage treatments, 200 g soil from net bag was taken for the determination of soil SOC, MBC and DOC. Content of cellulose, hemicellulose and lignin in straw were determined following Van’s method30, using a SLQ-6A semi-automatic crude fiber analyzer (Shanghai Fiber Testing Instrument Co., Ltd.).The following formula was used to calculate decomposition rate of straw and its components. M0 is the initial straw or cellulose (hemicellulose, lignin) mass, g, and Mt is the straw or cellulose (hemicellulose, lignin) mass at time t, g.$$mathrm{Decomposition ; proportion }left({%}right)= frac{{M}_{ 0}-{ M}_{ t}}{{M , }_{0}}times 100.$$
(1)
The following formula was used to calculate the straw carbon release proportion. C0 is the initial straw carbon content, g, Ct is the straw carbon content at time t, g.$$mathrm{Straw ; carbon ; release ; proportion }left({%}right)= frac{{C}_{ 0} -{ C }_{t}}{{C }_{0}}times 100.$$
(2)
The following formula was used to calculate the straw and its components decomposition rate. M365 is the quality of straw or cellulose (hemicellulose, lignin) mass on the 365th day, mg day−1.$$mathrm{Decomposition ; rate }left(mathrm{mg }{mathrm{day}}^{-1}right)= frac{{M}_{ 0 }- {M}_{365}}{365}.$$
(3)
The relationship of the straw decomposition proportion (%) changes over time was fitted as follows:$${y}_{t} = a+btimes exp left(-ktright),$$
(4)
where yt is the proportion of the straw decomposition proportion at time t, %; t is the decomposition time of straw; k is the decomposition rate constant calculated using the least-squares method; a and b are constants.SOC concentrations (g kg−1) was determined using the K2Cr2O7–H2SO4 digestion method31. Soil MBC content was determined using the Chloroform fumigation extraction method32. Two fresh soil samples were weighed, and then one of them was placed in a vacuum dryer with chloroform added, and pumped until the chloroform boiled violently, and after a period of time, the dryer cover was opened, the container containing chloroform removed, and the lid replaced. Another portion of soil was placed in a vacuum dryer without chloroform as a control. Then, 20 g each of fumigated and unfumigated soil samples were weighed, 50 mL 0.5 mg L−1 K2SO4 was added, extracted by vibration for 0.5 h, filtrate was pumped by 0.45 μm organic filter membrane, and then the filtrate was directly analyzed and detected using a TOC organic carbon analyzer. Based on the difference of organic C content between fumigated and unfumigated soil extracts, the microbial biomass carbon was obtained by multiplying the coefficient by 2.64. For the determination of soil DOC content, we used a slightly modified method of Jones33 and Hu Haiqing34. We made a leaching solution with 0.5 mol L K2SO4, weighed 10 g over 2 mm sieve of air dried soil, added the soil to the leaching solution to create a soil mass ratio of 2.5:1, and then applied a shock temperature for 1 h (220 r min−1). Then, after filtering, the filtrate was centrifuged for 20 min (3800 r min−1), filtered with a 0.45 μm organic membrane, and the filtrate subjected to TOC organic carbon analysis meter tests.Data analysisIn this experiment, Excel 2016 (Microsoft Corporation, New Mexico, USA) software was used to collate and analyze the data, and SPSS 19.0 (SPSS Inc., Chicago, Illinois, USA) statistical software was used to conduct variance analysis, LSD multiple analysis comparison and nonlinear regression analysis on the data. Duncan’s multiple range test was used to compare the treatment means at a 95% confidence level. Graphs were drawn using Origin 9.0 (Originlab, Northampton, USA). More

We processed DNB imagery from three key regions per the historical record of mariner sightings2—the northwest Indian Ocean (5° S–20° N, 40–70° E) and Indonesian waters surrounding Java (15° S–0°, 100–115° E) and the Banda Sea (11–1° S, 120–135° E). Our search window spanned 2012–2021, during the periods December–March and July–September corresponding to the peak modes of ship sightings. Data processing and detection criteria are described in “Methods” section.Our search yielded 12 DNB-detected events (listed in Table 1) whose properties met the strict criteria for milky seas. Physically unexplainable in terms of thermal emissions (which would require scene temperatures exceeding 600 K), uncorrelated with clouds/airglow, invisible during the day, and persistent over multiple consecutive nights, these luminous bodies drifted and evolved in ways that were consistent with the analyzed ocean surface currents. The start and end dates of detection were in many cases bound by the observable periods as defined by the lunar cycle. Here, we highlight three exemplary cases, with additional details for all cases summarized in Supplementary Discussion 2.Table 1 Day/Night Band detected milky sea events identified in this study.Full size tableSocotra, July/August 2013On 31 July 2013, the Suomi NPP DNB detected a luminous body with well-defined boundaries (Fig. 2), located east of Socotra in the northwest Indian Ocean, at (14.0° N, 57.0° E). Uncorrelated with the observed cloud field (Fig. 2a–c), the body drifted northeast with the currents at ~ 0.44 m s−1, stretching and curving in a manner consistent with the analysed ocean-surface currents (Fig. 2d–f), which showed a clockwise-rotating eddy located to its south.Figure 2Three-night sequence over 2–4 August 2013 of a bioluminescent milky sea in the Arabian Sea for (a–c) DNB log10—scaled radiance imagery (W cm−2 sr−1), showing a ~ 9000 km2 luminous body persisting amidst the ephemeral cloud cover, and (d–f) a pan-out of HYCOM sea surface currents (magenta box in (d) corresponds to domain of (a–c), with approximate location of the luminous body noted) shown for comparison against the body’s observed structural evolution and drift.Full size imageWhereas DNB imagery showed only dark ocean during the daytime overpasses of the same location, these glowing waters persisted on successive nights over a two-week period. By 2 August the milky sea covered ~ 9000 km2 (involving roughly 5 × 1021 to 5 × 1022 luminous bacteria, per “Methods” section). The DNB lost sight of the milky sea on 14 August due to moonlight, and it was not seen again in the following moon-free period.Suomi NPP’s daytime chlorophyll-a (Chla) retrievals, a proxy for the amount of organic material in the surface waters, showed structural similarities to the milky sea, but were more widespread (Supplementary Fig. S1). Moreover, the most elevated regions of Chla ( > 1 mg m−3) occurred not directly atop, but adjacent to the most luminous waters—a recurring property among the cases documented in this research which may indicate regions of algal stress where (potentially luminous) bacteria would proliferate. On several nights, a faint signature of the luminous body was detectable beneath analyzed cloud cover; its light scattering upward through the clouds in a way similar to the behaviour of city lights in DNB imagery.Somali Sea, January 2018On 12 January 2018, both Suomi NPP and NOAA-20 captured a luminous structure offshore of southern Somalia. Over the next 5 days it stretched into a narrow filament that paralleled the Somali coast, mirroring the behaviour of other winter-mode Somali Sea cases described in Supplementary Discussion 2. Over 18–23 January, the luminous filament extended east/northeast, forming a comma-shape (Fig. 3a–c), with a sharply-defined southeastern edge and gradually fading brightness on its northwestern side. By 20 January, it spanned ~ 15,000 km2, suggesting involvement of roughly 8 × 1021 to 8 × 1022 bacteria. Its scale, shape, time, and location were similar to the Lima-sighted milky sea4,14, as well as to a subset of the surface reports in Supplementary Discussion 1.Figure 3Three-night sequence over 20–22 January 2018 of a bioluminescent milky sea in the Somali Sea for (a–c) DNB log10 – scaled radiance imagery (W cm−2 sr−1), showing a ~ 15,000 km2 luminous feature persistent amidst the variable cloud field. Focusing on 22 January, (d) VIIRS-derived night time SST (K; with cloud cover in black) at 2156Z, (e) daytime VIIRS-retrieved Chla (mg m−3) at 1007Z, and (f) pan-out of HYCOM sea surface currents at 2100Z with approximate location of DNB-observed luminous body.Full size imageComparing the luminous body to satellite retrievals of Sea Surface Temperature (SST; Fig. 3d) showed its eastern boundary aligned with the edge of an oceanic front, residing within relatively cool (298–299 K) waters that extended northeast from the Somali coastal upwelling zone. These cooler SSTs corresponded to elevated Chla values in the range of ~ 0.5–1.0 mg m−3 (Fig. 3e). As in the 2013 Socotra case, the area of elevated Chla was more extensive than the luminous region. Ocean surface currents (Fig. 3f) showed the body’s eastern boundary embedded within counter-clockwise flow and drifting north/northwest at ~ 0.8 m s−1.Java, July–September 2019The DNB detected a large milky sea in the east Indian Ocean, immediately south of Java, Indonesia in 2019. The event spanned two complete moon-free cycles (26 July–9 August, and 25 August–7 September). On the night of 25 July, the DNB detected a luminous anomaly south of Surakarta, Java, near 9.5° S, 111° E. The detection amidst moderate moonlight conditions suggested a particularly strong source of emission. Imagery on subsequent moonless nights confirmed that the initial detection was in fact part of a much larger milky sea, spanning ~ 100,000 km2—approximately the same size as Iceland. A milky sea of this scale suggests involvement of roughly 6 × 1022 to 6 × 1023 luminous bacteria, which would qualify as the largest event on record. Undetectable during the day, the contiguous feature reappeared in nightly imagery throughout the two observable moon-free periods (Fig. 4).Figure 4Day/night comparison of DNB log10—scaled radiance imagery (W cm−2 sr−1) of a bioluminescent milky sea near Java for the period 2–4 August 2019 for (a–c) daytime imagery, and (d–f) night time imagery. The amorphous luminous body, located immediately south of Java and detectable only at night, covered ~ 100,000 km2 of ocean surface. Bright patches seen over Java in (d–f) are city lights.Full size imageSituated within quiescent, low-shear waters (Supplementary Fig. S2) between counter-clockwise-spinning warm-core eddies to its southeast and southwest (Fig. 4), this massive milky sea rotated clockwise like a cog between gears, its centre near 9.0° S, 110.0° E. By 30 July, its northern boundary approached within 25 km of the Java coast, and a 500 km2 area of its core was so bright that certain infrared-detected cumulus clouds appeared in the DNB imagery as dark, attenuating objects in contrast to the glowing waters below them (Supplementary Fig. S3).The DNB radiances measured in the brightest areas of these luminous waters approached Crescent- to Quarter-Moon illumination levels. Based on scotopic vision sensitivity to bioluminescent emission (Supplementary Fig. S4) and direct comparisons against legacy OLS imagery (Supplementary Fig. S5), portions of this milky sea may have appeared visually bright to dark-adapted human vision—perhaps even attaining the classical snowfield effect described in the historical mariner accounts.After losing sight of the luminous body on 10 August due to moonlight contamination, the DNB recaptured it on 25 August and tracked it for 2 weeks thereafter, as described in Supplementary Discussion 2. The longevity of this event, which lasted for at least 45 nights, by far eclipsed all other cases encountered in this study—indicating that significant milky seas offer a reasonable time window for reaching them if a rapid-response team is on the ready.Satellite retrievals of ocean surface properties for the 2019 Java case (Fig. 5) showed cooler waters and elevated Chla along the Java coast. A stream of higher Chla ( > 2 mg m−3), embedded within a tongue of these cooler waters, extended southward and immediately east of the luminous body, following the flow of the south-eastern eddy (Fig. 5b). At this time, the luminous waters were confined to a narrow range of SST over 298 ± 1 K (~ 25 ± 1 °C) and moderate Chla over 1 ± 0.5 mg m−3, demarcated from the surrounding waters at abrupt boundaries defined by coastal upwelling to the north and the two eddies to the south.Figure 5Multi-parameter analysis of the 2019 Java milky sea on 2 August 2019 for (a) night time DNB log10—scaled radiance imagery (W cm−2 sr−1) at 1752Z, (b) pan-out of HYCOM sea surface currents valid at 1800Z (magenta box shows domain of (a), with approximate location of luminous body shaded), (c) VIIRS-retrieved SST (with cloud cover and land surfaces in black) valid at 1752Z, and (d) daytime VIIRS-retrieved Chla at 0554Z.Full size imageFigure 6 relates DNB radiance, SST, and Chla for 10 nights (27 July–5 August) of the 2019 Java case, centred on the luminous body. DNB radiances correlated positively with Chla over 0.5–1.5 mg m−3, and negatively with SST over 297–299 K. The brightest milky sea waters corresponded to asymptotic SST values of ~ 298 K and Chla of ~ 1.2 mg m−3, respectively, and notably, did not overlap with the strongest parts of the algal bloom. The SST values, hovering around 298 K for most DNB-detected cases in this study, may hold significance, as this temperature regime promotes rapid growth of V. harveyi and P. leiognathi over a wide range of ocean salinity values20.Figure 6Relationship between DNB-measured milky sea radiances and ocean surface fields for the 2019 Java case (27 July–5 August, centred on the luminous body). Radiance-specific distributions (i.e., for a given radiance level, each row sums to 100%) are shown as a function of (a) SST and (b) Chla. The DNB noise floor (where SNR = 1) is drawn as a horizontal dashed line.Full size image More

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