Pre-registration: ecology → P1 fraction, out-of-sample panel

Pre-registration of a prospective seven-species test of Perez’s strict-octave prediction λ_Prot/λ_DNA = log 2/log φ ≈ 1.4404 in basal land-plant mitochondria. The hypothesis under test is that the strict-octave regime is the ancestral state of land-plant mitochondrial DNA–proteome coupling, preserved in bryophytes and lycophytes and progressively lost across the radiation of seed plants. Two prior anchor observations (Physcomitrium patens, Selaginella moellendorffii; both sub-1% from the predicted ratio) motivate the test; this pre-registration extends to seven additional species spanning mosses, liverworts, hornworts, and lycophytes. The decision rule is fixed in advance: ≥4/7 sub-1% matches confirms the ancestral-state hypothesis, ≤1/7 refutes it, intermediate values are inconclusive. Six methodological cross-checks (randomised-encoding null, nuclear-genome control, length-filter robustness, codon-bias correction, bootstrap CI, non-basal outgroup) and three robustness conditions are pre-specified. The analysis pipeline has not been executed at the time of pre-registration. Timestamped 2026-05-26.Pre-registration and execution of an out-of-sample test of the c ↔ f_P1 regression bridging the dynamical Lyapunov invariant and the static P1/P2 partition balance of the Master Code of Biology framework. Salinibacter ruber, a halophilic bacterium with reported disorder factor c = 0.7369, was selected as a discriminating test case. The pre-registration fixed the predicted f_P1 = 56.59% with band [54.59, 58.59] before any computational measurement was performed. Subsequent UniProt Swiss-Prot 2026_01 measurement returned 54.75% (strain M31) or 56.29% (species), both within the pre-registered band. The result supports the linearity of the c ↔ f_P1 relationship under composition-based predictions. Pre-registration timestamped 2026-05-26T14:58:38Z; measurement performed in the same session, after the pre-registration was finalised. Pre-registration and execution of an ecology-based out-of-sample test of f_P1 partition balance across a 16-organism panel spanning four ecological strategies (halophiles, hyperthermophiles, mesophiles, psychrophiles) and two domains (bacteria, archaea). Anchored predictions derived from in-sample organisms were stated in advance: halophiles f_P1 ≈ 58 ± 2%, hyperthermophiles ≈ 45 ± 2%, mesophiles ≈ 51 ± 2%, with psychrophiles left as exploratory. Of ten anchored organisms tested, nine fell within the pre-registered band. The single outlier was Chromohalobacter salexigens, a halophilic bacterium using a compatible-solute strategy distinct from the salt-in strategy of the haloarchaeal anchors; this is biologically interpretable rather than a generic failure of the ecology framing. Psychrophiles clustered at 50.7–52.8%, similar to mesophiles — a new exploratory finding. Pre-registration timestamped 2026-05-26T15:30:00Z; measurement performed in the same session, after the pre-registration was finalised.针对陆生植物基部类群线粒体中Perez严格八度预测λ_Prot/λ_DNA = log₂/logφ ≈1.4404的前瞻性7物种测试的预注册（Pre-registration）。本测试所检验的假说为：严格八度机制是陆生植物线粒体DNA-蛋白质组耦合的祖先状态，该状态在苔藓植物（bryophytes）和石松类植物（lycophytes）中得以保留，并在种子植物的辐射演化过程中逐渐丢失。两项先前的锚定观测（小立碗藓（Physcomitrium patens）和卷柏（Selaginella moellendorffii），二者与预测比值的偏差均小于1%）为本测试提供了依据；本次预注册将测试范围扩展至另外7个物种，涵盖苔藓、地钱（liverworts）、角苔（hornworts）和石松类植物。决策规则已预先设定：若7个物种中有≥4个与预测比值的偏差小于1%，则证实该祖先状态假说；若≤1个符合，则推翻该假说；介于两者之间的结果则无定论。已预先设定6项方法学交叉检验（随机编码零假设、核基因组对照、长度过滤稳健性检验、密码子偏好性校正、bootstrap置信区间（bootstrap CI）、非基部类群外类群）以及3项稳健性条件。本预注册完成时，分析流程尚未执行。预注册时间戳为2026-05-26。 针对生物学主密码（Master Code of Biology）框架中连接动态李雅普诺夫不变量与静态P1/P2划分平衡的c ↔ f_P1回归的样本外测试的预注册与执行。选取已报道无序因子c=0.7369的嗜盐细菌红嗜盐碱菌（Salinibacter ruber）作为区分性测试样本。在开展任何计算测量之前，本次预注册已确定预测的f_P1为56.59%，置信区间为[54.59, 58.59]。后续基于UniProt Swiss-Prot 2026_01版本的测量结果显示，菌株M31的f_P1为54.75%，该物种整体的f_P1为56.29%，二者均落在预注册设定的置信区间内。该结果支持基于组分预测时，c与f_P1之间存在线性关系。预注册时间戳为2026-05-26T14:58:38Z；测量于同一批次中完成，且在预注册最终确定之后进行。 针对涵盖4种生态策略（嗜盐菌、超嗜热菌、嗜温菌、嗜冷菌）和2个域（细菌域（bacteria）、古菌域（archaea））的16物种测试组的f_P1划分平衡的基于生态学的样本外测试的预注册与执行。基于样本内物种得到的锚定预测已预先声明：嗜盐菌f_P1≈58±2%，超嗜热菌≈45±2%，嗜温菌≈51±2%，嗜冷菌则作为探索性测试对象。在测试的10个锚定物种中，有9个落在预注册设定的区间内。唯一的异常值是盐单胞菌（Chromohalobacter salexigens），这是一种采用相容性溶质策略的嗜盐细菌，与锚定的盐古菌采用的盐入策略截然不同；该异常值具有生物学解释性，并非生态学框架的一般性失效。嗜冷菌的f_P1值聚集在50.7%~52.8%之间，与嗜温菌相近——这是一项新的探索性发现。预注册时间戳为2026-05-26T15:30:00Z；测量于同一批次中完成，且在预注册最终确定之后进行。